A reagent commonly used in biochemical studies as a protective agent to prevent the oxidation of SH (thiol) groups and for reducing disulphides to dithiols.
Materials that add an electron to an element or compound, that is, decrease the positiveness of its valence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed)
Chemical agents that react with SH groups. This is a chemically diverse group that is used for a variety of purposes. Among these are enzyme inhibition, enzyme reactivation or protection, and labelling.
Compounds containing the -SH radical.
Chemical groups containing the covalent disulfide bonds -S-S-. The sulfur atoms can be bound to inorganic or organic moieties.
A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).
A thiol-containing non-essential amino acid that is oxidized to form CYSTINE.
A sulfhydryl reagent that is widely used in experimental biochemical studies.
The rate dynamics in chemical or physical systems.
A standard reagent for the determination of reactive sulfhydryl groups by absorbance measurements. It is used primarily for the determination of sulfhydryl and disulfide groups in proteins. The color produced is due to the formation of a thio anion, 3-carboxyl-4-nitrothiophenolate.
A sulfhydryl reagent which oxidizes sulfhydryl groups to the disulfide form. It is a radiation-sensitizing agent of anoxic bacterial and mammalian cells.
A tripeptide with many roles in cells. It conjugates to drugs to make them more soluble for excretion, is a cofactor for some enzymes, is involved in protein disulfide bond rearrangement and reduces peroxides.
Benzoic acid esters or salts substituted with one or more iodine atoms.
Compounds which restore enzymatic activity by removing an inhibitory group bound to the reactive site of the enzyme.
The sum of the weight of all the atoms in a molecule.
The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.
An alkylating sulfhydryl reagent. Its actions are similar to those of iodoacetate.
Hydrogen-donating proteins that participates in a variety of biochemical reactions including ribonucleotide reduction and reduction of PEROXIREDOXINS. Thioredoxin is oxidized from a dithiol to a disulfide when acting as a reducing cofactor. The disulfide form is then reduced by NADPH in a reaction catalyzed by THIOREDOXIN REDUCTASE.
Chloride and mercury-containing derivatives of benzoic acid.
Hydroxylated benzoic acid derivatives that contain mercury. Some of these are used as sulfhydryl reagents in biochemical studies.
An ethylmercury-sulfidobenzoate that has been used as a preservative in VACCINES; ANTIVENINS; and OINTMENTS. It was formerly used as a topical antiseptic. It degrades to ethylmercury and thiosalicylate.
A compound that, along with its isomer, Cleland's reagent (DITHIOTHREITOL), is used for the protection of sulfhydryl groups against oxidation to disulfides and for the reduction of disulfides to sulfhydryl groups.
A chelating agent that sequesters a variety of polyvalent cations such as CALCIUM. It is used in pharmaceutical manufacturing and as a food additive.
A GLUTATHIONE dimer formed by a disulfide bond between the cysteine sulfhydryl side chains during the course of being oxidized.
Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.
A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.
A strong oxidizing agent used in aqueous solution as a ripening agent, bleach, and topical anti-infective. It is relatively unstable and solutions deteriorate over time unless stabilized by the addition of acetanilide or similar organic materials.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.
An anti-gas warfare agent that is effective against Lewisite (dichloro(2-chlorovinyl)arsine) and formerly known as British Anti-Lewisite or BAL. It acts as a chelating agent and is used in the treatment of arsenic, gold, and other heavy metal poisoning.
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
An organic mercurial used as a sulfhydryl reagent.
A sulfur-containing alkyl thionitrite that is one of the NITRIC OXIDE DONORS.
Electron-accepting molecules in chemical reactions in which electrons are transferred from one molecule to another (OXIDATION-REDUCTION).
A radiation-protective agent that interferes with sulfhydryl enzymes. It may also protect against carbon tetrachloride liver damage.
Iodinated derivatives of acetic acid. Iodoacetates are commonly used as alkylating sulfhydryl reagents and enzyme inhibitors in biochemical research.
The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).
Inorganic derivatives of phosphorus trihydroxide (P(OH)3) and its tautomeric form dihydroxyphosphine oxide (HP=O(OH)2). Note that organic derivatives of phosphonic acids are listed under are ORGANOPHOSPHONATES.
The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.
Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.
An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter.
Presence of warmth or heat or a temperature notably higher than an accustomed norm.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Separation technique in which the stationary phase consists of ion exchange resins. The resins contain loosely held small ions that easily exchange places with other small ions of like charge present in solutions washed over the resins.
A serine endopeptidase that is formed from TRYPSINOGEN in the pancreas. It is converted into its active form by ENTEROPEPTIDASE in the small intestine. It catalyzes hydrolysis of the carboxyl group of either arginine or lysine. EC 3.4.21.4.
The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.
A covalently linked dimeric nonessential amino acid formed by the oxidation of CYSTEINE. Two molecules of cysteine are joined together by a disulfide bridge to form cystine.
Nicotinamide adenine dinucleotide phosphate. A coenzyme composed of ribosylnicotinamide 5'-phosphate (NMN) coupled by pyrophosphate linkage to the 5'-phosphate adenosine 2',5'-bisphosphate. It serves as an electron carrier in a number of reactions, being alternately oxidized (NADP+) and reduced (NADPH). (Dorland, 27th ed)
Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.
Positively charged atoms, radicals or groups of atoms with a valence of plus 2, which travel to the cathode or negative pole during electrolysis.
A chromatographic technique that utilizes the ability of biological molecules to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
A metallic element that has the atomic symbol Mg, atomic number 12, and atomic weight 24.31. It is important for the activity of many enzymes, especially those involved in OXIDATIVE PHOSPHORYLATION.
A basic element found in nearly all organized tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
A subtype of thioredoxins found primarily in CHLOROPLASTS.
Analogs of those substrates or compounds which bind naturally at the active sites of proteins, enzymes, antibodies, steroids, or physiological receptors. These analogs form a stable covalent bond at the binding site, thereby acting as inhibitors of the proteins or steroids.
The covalent bonding of an alkyl group to an organic compound. It can occur by a simple addition reaction or by substitution of another functional group.
Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby forming bridges between side chains of amino acids in proteins; the locations of naturally reactive areas within proteins can thereby be identified; may also be used for other macromolecules, like glycoproteins, nucleic acids, or other.
A class of enzymes that catalyze geometric or structural changes within a molecule to form a single product. The reactions do not involve a net change in the concentrations of compounds other than the substrate and the product.(from Dorland, 28th ed) EC 5.
A basic science concerned with the composition, structure, and properties of matter; and the reactions that occur between substances and the associated energy exchange.
A group of compounds that contain a bivalent O-O group, i.e., the oxygen atoms are univalent. They can either be inorganic or organic in nature. Such compounds release atomic (nascent) oxygen readily. Thus they are strong oxidizing agents and fire hazards when in contact with combustible materials, especially under high-temperature conditions. The chief industrial uses of peroxides are as oxidizing agents, bleaching agents, and initiators of polymerization. (From Hawley's Condensed Chemical Dictionary, 11th ed)
Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.
Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.
The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat.
The composition, conformation, and properties of atoms and molecules, and their reaction and interaction processes.
Sulfur-sulfur bond isomerases that catalyze the rearrangement of disulfide bonds within proteins during folding. Specific protein disulfide-isomerase isoenzymes also occur as subunits of PROCOLLAGEN-PROLINE DIOXYGENASE.
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
The class of all enzymes catalyzing oxidoreduction reactions. The substrate that is oxidized is regarded as a hydrogen donor. The systematic name is based on donor:acceptor oxidoreductase. The recommended name will be dehydrogenase, wherever this is possible; as an alternative, reductase can be used. Oxidase is only used in cases where O2 is the acceptor. (Enzyme Nomenclature, 1992, p9)
Proteins prepared by recombinant DNA technology.
A toxic thiol mercury salt formerly used as a diuretic. It inhibits various biochemical functions, especially in mitochondria, and is used to study those functions.
Purifying or cleansing agents, usually salts of long-chain aliphatic bases or acids, that exert cleansing (oil-dissolving) and antimicrobial effects through a surface action that depends on possessing both hydrophilic and hydrophobic properties.
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point.
Genetically identical individuals developed from brother and sister matings which have been carried out for twenty or more generations or by parent x offspring matings carried out with certain restrictions. This also includes animals with a long history of closed colony breeding.
A coenzyme composed of ribosylnicotinamide 5'-diphosphate coupled to adenosine 5'-phosphate by pyrophosphate linkage. It is found widely in nature and is involved in numerous enzymatic reactions in which it serves as an electron carrier by being alternately oxidized (NAD+) and reduced (NADH). (Dorland, 27th ed)
Disruption of the non-covalent bonds and/or disulfide bonds responsible for maintaining the three-dimensional shape and activity of the native protein.
A diverse group of agents, with unique chemical structures and biochemical requirements, which generate NITRIC OXIDE. These compounds have been used in the treatment of cardiovascular diseases and the management of acute myocardial infarction, acute and chronic congestive heart failure, and surgical control of blood pressure. (Adv Pharmacol 1995;34:361-81)
The ability of a substance to be dissolved, i.e. to form a solution with another substance. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts.
The anterior portion of the spermatozoon (SPERMATOZOA) that contains mainly the nucleus with highly compact CHROMATIN material.
Multicellular, eukaryotic life forms of kingdom Plantae (sensu lato), comprising the VIRIDIPLANTAE; RHODOPHYTA; and GLAUCOPHYTA; all of which acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations.
A cytotoxic sulfhydryl reagent that inhibits several subcellular metabolic systems and is used as a tool in cellular physiology.

Regulation of 2-carboxy-D-arabinitol 1-phosphate phosphatase: activation by glutathione and interaction with thiol reagents. (1/2188)

2-Carboxy-D-arabinitol 1-phosphate (CA1P) phosphatase de- grades CA1P, an inhibitor associated with the regulation of ribulose bisphosphate carboxylase/oxygenase in numerous plant species. CA1P phosphatase purified from Phaseolus vulgaris was partially inactivated by oxidizing conditions during dialysis in air-equilibrated buffer. Phosphatase activity could then be stimulated 1.3-fold by dithiothreitol and also by addition of reduced thioredoxin from Escherichia coli. These effects were enhanced synergistically by the positive effector, fructose 1, 6-bisphosphate (FBP). Most notably, CA1P phosphatase activity was stimulated up to 35-fold by glutathione, and was sensitive to the ratio of reduced (GSH) to oxidized (GSSG) forms. At concentrations of glutathione approximating measured levels in chloroplasts of P. vulgaris (5 mM total S), CA1P phosphatase exhibited >20-fold stimulation by a change in the redox status of glutathione from 60 to 100% GSH. This stimulation was augmented further by reduced E. coli thioredoxin. In contrast, FBP, which activates CA1P phosphatase under reducing conditions, was strongly inhibitory in the presence of GSSG. We propose that glutathione may have an appreciable role in the light/dark regulation of CA1P phosphatase in vivo. A model for the reversible activation of CA1P phosphatase by GSH was derived based upon the various responses of the enzyme's activity to a range of thiol reagents including N-ethylmaleimide, 5, 5'-dithiobis-(2-nitrobenzoic acid) and arsenite. These data indicate that the bean enzyme contains two physically distinct sets of thiol groups that are critical to its redox regulation.  (+info)

In vivo formation of Cu,Zn superoxide dismutase disulfide bond in Escherichia coli. (2/2188)

We have found that the in vivo folding of periplasmic Escherichia coli Cu,Zn superoxide dismutase is assisted by DsbA, which catalyzes the efficient formation of its single disulfide bond, whose integrity is essential to ensure full catalytic activity to the enzyme. In line with these findings, we also report that the production of recombinant Xenopus laevis Cu,Zn superoxide dismutase is enhanced when the enzyme is exported in the periplasmic space or is expressed in thioredoxin reductase mutant strains. Our data show that inefficient disulfide bond oxidation in the bacterial cytoplasm inhibits Cu,Zn superoxide dismutase folding in this cellular compartment.  (+info)

An intact sperm nuclear matrix may be necessary for the mouse paternal genome to participate in embryonic development. (3/2188)

We have been interested in determining the minimally required elements in the sperm head that are necessary in order for the paternal genome to participate in embryogenesis. We used an ionic detergent, mixed alkyltrimethylammonium bromide (ATAB), plus dithiothreitol (DTT) to remove the acrosome and almost all of the perinuclear theca, leaving only the sperm nucleus morphologically intact. We also tested the stability of the sperm nuclear matrix by the ability to form nuclear halos. Sperm nuclei washed in freshly prepared 0.5% ATAB + 2 mM DTT completely decondensed when extracted with salt, but nuclei washed in the same buffer that was 1 wk old, and then extracted with salt, produced nuclear halos, indicating stable nuclear matrices. When we treated sperm heads with freshly prepared ATAB+DTT and injected them into oocytes, none of the oocytes developed into live offspring. In contrast, sperm heads treated in the same way but with 1-wk-old ATAB+DTT solution could support development of about 30% of the oocytes to live offspring. Electron microscopy demonstrated that most of the perinuclear theca had been removed in both cases. These data suggest that at least in the mouse, the only component of the spermatozoa that is crucial for participation in embryologic development is the sperm nucleus with a stable nuclear matrix.  (+info)

A novel trans-complementation assay suggests full mammalian oocyte activation is coordinately initiated by multiple, submembrane sperm components. (4/2188)

To initiate normal embryonic development, an egg must receive a signal to become activated at fertilization. We here report that the ability of demembranated sperm heads to activate is abolished after incubation over the range 20-44 degreesC and is sensitive to reducing agents. On the basis of this observation, we have developed a microinjection-based, trans-complementation assay in order to dissect the heat-inactivated sperm-borne oocyte-activating factor(s) (SOAF). We demonstrate that the failure of heat-inactivated sperm heads to activate an egg is rescued by coinjection with dithiothreitol-solubilized SOAF from demembranated sperm heads. The solubilized SOAF (SOAFs) is trypsin sensitive and is liberated from demembranated heads in a temperature-dependent manner that inversely correlates with the ability of sperm heads to activate. This argues that SOAFs is a proteinaceous molecular species required to initiate activation. Injection of oocytes with mouse or hamster sperm cytosolic factors, but not SOAFs alone, induced resumption of meiosis, further suggesting that these cytosolic factors and SOAF are distinct. Collectively, these data strongly suggest that full mammalian oocyte activation is initiated by the coordinated action of one or more heat-sensitive protein constituents of the perinuclear matrix and at least one heat-stable submembrane component.  (+info)

Transforming growth factor-beta induces formation of a dithiothreitol-resistant type I/Type II receptor complex in live cells. (5/2188)

Transforming growth factor-beta (TGF-beta) binds to and signals via two serine-threonine kinase receptors, the type I (TbetaRI) and type II (TbetaRII) receptors. We have used different and complementary techniques to study the physical nature and ligand dependence of the complex formed by TbetaRI and TbetaRII. Velocity centrifugation of endogenous receptors suggests that ligand-bound TbetaRI and TbetaRII form a heteromeric complex that is most likely a heterotetramer. Antibody-mediated immunofluorescence co-patching of epitope-tagged receptors provides the first evidence in live cells that TbetaRI. TbetaRII complex formation occurs at a low but measurable degree in the absence of ligand, increasing significantly after TGF-beta binding. In addition, we demonstrate that pretreatment of cells with dithiothreitol, which inhibits the binding of TGF-beta to TbetaRI, does not prevent formation of the TbetaRI.TbetaRII complex, but increases its sensitivity to detergent and prevents TGF-beta-activated TbetaRI from phosphorylating Smad3 in vitro. This indicates that either a specific conformation of the TbetaRI. TbetaRII complex, disrupted by dithiothreitol, or direct binding of TGF-beta to TbetaRI is required for signaling.  (+info)

Gamma-Actinin, a new regulatory protein from rabbit skeletal muscle. I. Purification and characterization. (6/2188)

A new regulatory protein which we have designated as gamma-actinin has been isolated from native thin filaments of rabbit skeletal muscle. Depolymerized native thin filaments were fractionated by salting out with ammonium sulfate, and the precipitates obtained at 40--60% ammonium sulfate saturation were further subjected to DEAE-Sephadex and Sephadex G-200 column chromatography. The purified gamma-actinin was shown to have a chain weight of 35,000 daltons and had a strong inhibitory action on the polymerization of G-actin. The results of amino acid analysis indicated a unique amino acid composition of gamma-actinin as compared with other structural proteins of muscle. Non-polar and neutral amino acid residues were abundant. One cysteine residue was contained per one molecule of gamma-actinin and played a critical role in the maintenance of the inhibitory activity. Pelleting of gamma-actinin with F-actin showed that gamma-actinin binds to F-action.  (+info)

Phospholipid hydroperoxide cysteine peroxidase activity of human serum albumin. (7/2188)

Human serum albumin (HSA) reduced the phospholipid hydroperoxide, 1-palmitoyl-2-(13-hydroperoxy-cis-9, trans-11-octadecadienoyl)-l-3-phosphatidylcholine (PLPC-OOH) to the corresponding hydroxy-derivative with a high apparent affinity (Km=9. 23+/-0.95 microM). Removal of bound lipid during purification increased this activity. At physiological concentration, HSA reduced the phospholipid hydroperoxide in the absence of a cofactor. However, in the presence of a cofactor (reductant), the rate of the reaction was increased. All of the major aminothiols in plasma could act as reductants, the best being the most abundant, cysteine (Km=600+/-80 microM). For every nanomole of PLPC-OOH reduced by HSA, 1.26 nmol of cystine was formed, indicating a reaction stoichiometry of 1 mol PLPC-OOH to 2 mol cysteine. We used chemical modification to determine which amino acid residues on HSA were responsible for the activity. Oxidation of thiol group(s) by N-ethylmaleimide led to a reduction in the rate of activity, whereas reduction of thiols by either dithiothreitol or the angiotensin-converting enzyme inhibitor, captopril, increased the activity. Both N-ethylmaleimide-modified HSA and dithiothreitol-treated HSA exhibited increased apparent affinities for PLPC-OOH. For a range of preparations of albumin with different modifications, the activity on PLPC-OOH was dependent on the amount of free thiol groups on the albumin (correlation coefficient=0.91). Patients with lowered albumin concentrations after septic shock showed lowered total plasma thiol concentrations and decreased phospholipid hydroperoxide cysteine peroxidase (PHCPx) activities. These results therefore show for the first time that HSA exhibits PHCPx activity, and that the majority of the activity depends on the presence of reduced thiol group(s) on the albumin.  (+info)

Respiratory chain strongly oxidizes the CXXC motif of DsbB in the Escherichia coli disulfide bond formation pathway. (8/2188)

Escherichia coli DsbB has four essential cysteine residues, among which Cys41 and Cys44 form a CXXC redox active site motif and the Cys104-Cys130 disulfide bond oxidizes the active site cysteines of DsbA, the disulfide bond formation factor in the periplasm. Functional respiratory chain is required for the cell to keep DsbA oxidized. In this study, we characterized the roles of essential cysteines of DsbB in the coupling with the respiratory chain. Cys104 was found to form the inactive complex with DsbA under respiration-defective conditions. While DsbB, under normal aerobic conditions, is in the oxidized state, having two intramolecular disulfide bonds, oxidation of Cys104 and Cys130 requires the presence of Cys41-Cys44. Remarkably, the Cys41-Cys44 disulfide bond is refractory to reduction by a high concentration of dithiothreitol, unless the membrane is solubilized with a detergent. This reductant resistance requires both the respiratory function and oxygen, since Cys41-Cys44 became sensitive to the reducing agent when membrane was prepared from quinone- or heme-depleted cells or when a membrane sample was deaerated. Thus, the Cys41-Val-Leu-Cys44 motif of DsbB is kept both strongly oxidized and strongly oxidizing when DsbB is integrated into the membrane with the normal set of respiratory components.  (+info)

Mice were killed and intestines were removed and placed in ice-cold HBSS + 1% heat inactivated FBS + 0.1% Pen/Strep (complete HBSS). After removal of residual mesenteric fat tissue, Peyers patches were carefully excised and the intestine was flushed two times with 10 ml complete HBSS. After flushing, the intestine was opened longitudinally and cut into ∼1-cm pieces. The cut pieces were washed two times at 37°C with 30 ml complete HBSS for 5 min each on a magnetic stir plate (600 rpm), followed by incubation with 50 ml 1-mM DTT for 15 min at 37°C with rotation. After DTT treatment, the tissue was then incubated twice with 30 ml 1-mM EDTA at 37°C with rotation. After each incubation, the epithelial cell layer containing the intraepithelial lymphocytes was removed by aspiration. After the last EDTA incubation, the pieces were washed three times in complete HBSS and placed in 30-ml digestion solution (complete HBSS containing Type III Collagenase and DNase I; Worthington). Digestion was ...
PlusOne Dithiothreitol from GE Healthcare, formerly Amersham Biosciences,Dithiothreitol, 1 g. High quality kits and reagents for convenient sample preparation that give consistent and reliable 2-D results.Effectively disrupt cells and tissues.Clean up samples from contaminants.Efficiently dialyze small sample volumes. Accurately quantitate protein for first-dimension is,biological,biology supply,biology supplies,biology product
CAS: 3483-12-3 Fórmula molecular: C4H10O2S2 Molecular Weight (g/mol): 154.24 Número MDL: MFCD00004877 InChI Key: VHJLVAABSRFDPM-IMJSIDKUSA-N Sinónimo: dithiothreitol, dl-1,4-dithiothreitol, dl-dithiothreitol, 1,4-dithio-dl-threitol, d-1,4-dithiothreitol, d-dtt, 2s,3s-1,4-dimercaptobutane-2,3-diol, threo-1,4-dimercapto-2,3-butanediol, 1,4-dithiothreitol, dtt PubChem CID: 446094 ChEBI: CHEBI:42170 IUPAC Name: (2S,3S)-1,4-bis(sulfanyl)butane-2,3-diol SMILES: C(C(C(CS)O)O)S 250ML DL-1,4-Dithiothreitol, for biochemistry, 1Msolution in water ...
Preparation of CYP3A4 Apo-Protein. Removal of the heme from CYP3A4 and preparation of the apo-protein was performed by treatment with H2O2 (Uvarov et al., 1990; Pikuleva et al., 1992). We implemented very mild conditions of treatment to prevent any peroxidative damage of the protein. In brief, CYP3A4 was diluted to 50 μM in 100 mM sodium phosphate buffer, pH 7.2, 20% glycerol (buffer A). Catalase was added directly to this solution to a final concentration of 1.7 units/ml. The sample was then dialyzed in a Spectra/Por Type I molecular weight cut-off 6000-8000, 10-cm flat width dialysis bag (Spectrum Laboratories, Rancho Dominguez, CA) against 50 volumes of buffer A containing 100 mM H2O2. Dialysis was performed at 4°C with continuous stirring for 24 h. The sample was removed and was further dialyzed against 50 volumes of 100 mM 4-morpholinepropanesulfonic acid, pH 7.4, 10% glycerol, 1 mM EDTA, 0.2 mM dithiothreitol. Protein concentration was determined using the Bradford protein assay kit ...
GFP-VHH:GFP complex is stable up to 80 °C, 1 mM DTT, 3 M Guanidinium•HCl, 8 M Urea, 2 M NaCl, 2 % Nonidet P40 Substitute, 1 % SDS, 1 % Triton X-100, 3 % ...
Human RAD51 from Invitrogen for Western Blot and Control applications. Supplied as 20 µg purified protein (1 mg/mL) in 20mM tris HCl with 100mM KCl, 10% glycerol, 1mM DTT, 0.5mM EDTA and no preservative; pH 8.
2 mM EDTA5 mM DTT1% Triton-XYeast protease inhibitor cocktail and phosphatase inhibitor cocktail (added fresh to the buffer B ...
I need to prepare the solution (50 mM Tris-HCl, pH 8.0, 500 mM KCl, 2 mM DTT, 1 mM EDTA, 1 mM orthovanadate, and 10% glycerol) Do you have some protocol or sell the prepared solution?. ...
Gentaur molecular products has all kinds of products like :search , Biotium \ DTT \ 91050 for more molecular products just contact us
The effect of a thiol, dithiothreitol (DTT), on DNA radiolysis at cryogenic temperatures was investigated by the EPR method. Dithiothreitol repairs up to 30% of DNA radicals, but an increase in the thiol concentration above 10 mM, even up to 1000 mM, does not further enhance the overall level of the repair process. The transfers of paramagnetic centres from both the guanine radical cation. and the allyl radical of thymine, to thiol were observed, while products generated via the reductive pathway remained unaffected. (C) 2002 Elsevier Science Ltd. All rights reserved.. ...
Effect of DTT on prolyl oligopeptidase levels in plasma from healthy and RR-MS patients. POP activity in plasma from healthy controls (white bar) and RR-MS pati
A compound that, along with its isomer, Clelands reagent (DITHIOTHREITOL), is used for the protection of sulfhydryl groups against oxidation to disulfides and for the reduction of disulfides to sulfhydryl groups ...
TY - JOUR. T1 - Insulin-dependent intermolecular subunit communication between isolated αβ heterodimeric insulin receptor complexes. AU - Sweet, L. J.. AU - Morrison, B. D.. AU - Wilden, P. A.. AU - Pessin, J. E.. PY - 1987/12/1. Y1 - 1987/12/1. N2 - The dissociation of the purified human placental α2β2 heterotetrameric insulin receptor complex into an αβ heterodimeric state was found to occur in a pH- and dithiothreitol (DTT)-dependent manner. Formation of the αβ heterodimeric complex, under conditions which preserved tracer insulin binding and protein kinase activities (pH 8.75 for 25 min followed by 2.0 mM DTT for 5 min) occurred with an approximate 50% efficiency. The resulting nondissociated α2β2 heterotetrameric complexes could then be separated effectively by Bio-Gel A-1.5 m gel filtration chromatography at neutral pH. The isolated DTT-treated but nondissociated α2β2 heterotetrameric complex was resistant to any further dissociation by a second round of DTT and alkaline pH ...
by fluorescence spectroscopy. Plant tissue was ground with a pestle and mortar in two volumes of extraction buffer (50mM Tris-HCl pH 8.0, 10mM EDTA, 10mM dithiothreitol, 18% glycerol) and filtered through Miracloth (Calbiochem). The filtrate was centrifuged to pellet debris and the supernatant collected. The supernatant was diluted ten fold with buffer and purified through a PD10 column (Pharmacia), which had been equilibrated in buffer. The protein content of the extracts was determined by the method of Bradford (Bradford, 1976) using a kit from BioRad Laboratories. Preparations were diluted with buffer to 80&g per ml and analyzed in a Kontron SFM25 scanning fluorometer ...
CD1B Recombinant Protein. CD1B protein solution (0.5mg/ml) containing 20mM Tris-HCl buffer (pH 8.0), 0.2M NaCl, 20% glycerol and 1mM DTT.
Supplier: SignalChem • Format: 50mM Tris-HCl, pH 7.5, 50mM NaCl, 10mM glutathione, 0.1mM EDTA, 0.25mM DTT, 0.1mM PMSF and 25% ...
MCF-7 cells (∼2.5 × 106) at 70-80% confluence in 15-cm plates were treated with 0.5 μm TSA in 0.1% ethanol or 0.1% ethanol as vehicle control for 24 h at 37°C. After washing with chilled DPBS, cells were harvested in DPBS, pelleted by centrifugation (1000 × g for 5 min at 4°C), and resuspended in 100 μl of high salt buffer [400 mm KCl, 20 mm Tris (pH 7.4), 2 mm DTT, and 20% (v/v) glycerol] containing a freshly added mixture of protease inhibitors (1 mm PMSF and 0.5 μg/ml each of leupeptin, pepstatin A, chymostatin, antitrypsin, and aprotinin). Whole cell extracts were prepared by three cycles of freezing (−80°C) and thawing (0°C), clarified by centrifugation at 15,000 × g for 15 min at 4°C, and the supernatants were stored at −80°C (13) . Protein concentration of each extract was determined using the Bio-Rad Protein Assay.. Each whole cell extract (500 μg protein) was suspended in 1 ml of buffer A [400 mm NaCl, 50 mm Tris acetate (pH 7.5), 1 mm DTT, 1% (v/v) Triton X-100, 1 mm ...
KD=7 nM. Caliper IC50 assay. The assay was performed using 384-well microtiter plates. Compounds were tested as 8-point dose responses. The assays were prepared by addition of 50 nL of compound solution in 90% DMSO directly into the empty plate. Subsequently, 4.5 μL of the enzyme solution (50 mM HEPES pH 7.5, 1 mM DTT, 0.02% Tween20, 0.02% BSA, 10 mM beta-glycerolphosphate, 10 μM sodium orthovanadate, 3 mM MgCl2 and 0.6 nM PAK1 (249-545) wt nonphos, produced in-house by expression in E. coli cells and purified by affinity chromatography) was added to each well and the resulting solution was pre-incubated at 30°C for 60 min, followed by addition of 4.5 μL of the peptide/ATP solution (50 mM HEPES pH 7.5, 1 mM DTT, 0.02% Tween20, 0.02% BSA, 10 mM beta-glycerolphosphate, 10 μM sodium orthovanadate, 3 mM MgCl2, 400 μM ATP and 4 μM peptide (5-Fluo-Ahx-AKRRRLSSLRA-COOH, Biosyntan GmbH). After 60 min incubation at 30°C, reactions were terminated by addition of 16 μL per well of the stop ...
sample_1: HypC, [U-95% 15N], 1 mM; potassium phosphate 30 mM; KCl 30 mM; dithiothreitol 25 uM; sodium azide 0.02%. sample_2: HypC, [U-95% 13C; U-95% 15N], 1 mM; potassium phosphate 30 mM; KCl 30 mM; dithiothreitol 25 uM; sodium azide 0.02%. sample_conditions_1: ionic strength: 0.06 M; pH: 7.0; pressure: 1 atm; temperature: 298 K ...
This product includes 3 vials: 1 vial of gene-specific cell lysate, 1 vial of control vector cell lysate, and 1 vial of loading buffer. Each lysate vial contains 0.1 mg lysate in 0.1 ml (1 mg/ml) of RIPA Buffer (50 mM Tris-HCl pH7.5, 250 mM NaCl, 5 mM EDTA, 50 mM NaF, 1% NP40). The loading buffer vial contains 0.5 ml 2X SDS Loading Buffer (125 mM Tris-Cl, pH6.8, 10% glycerol, 4% SDS, 0.002% Bromophenol blue, 100 mM DTT).. ...
Anterior Gradient Protein 3 Homolog Recombinant. AGR3 protein solution (1mg/ml) containing 20mM Tris-HCl buffer (pH8.0), 20% glycerol, 0.1M NaCl and 1mM DTT.
While I dont know any literature regarding this, the best thing that can be done from my experience, is to dissolve the DTT in high quality deionized water that has been degassed thoroughly prior to use. The biggest issue for DTT is going to be oxygen in your solvents. Next, the DTT is aliquoted into smaller quantities (generally enough for one days worth of reactions), and the aliquots are frozen at -20 or flash frozen using liquid nitrogen and stored at -80C. ...
* found in: Dithiothreitol, DeStreak Rehydration Solution, DeStreak Reagent, Reducing agents are frequently included in the sample solution to break any..
For this procedure, the cells are first swollen in a buffered hypo‐osmotic medium containing potassium chloride, magnesium acetate and dithiothreitol to render the plasma membrane more susceptible to subsequent homogenization
Los Angeles, CA (PRWEB) September 06, 2012 -- DTT announces a partnership with Sarku Japan®, the largest Japanese QSR operator in the US. After extensive
sample_1: BC022030, [U-15N; U-13C], 0.5 ± 0.1 mM; Bis-Tris 10 ± 0.5 mM; DTT 10 ± 0.5 mM; NaCl 100 ± 5 mM. sample_2: BC022030, [U-15N; U-13C], 0.5 ± 0.1 mM; BC022030 0.5 ± 0.1 mM; Bis-Tris 10 ± 0.5 mM; DTT 10 ± 0.5 mM; NaCl 100 ± 5 mM. conditions_1: pH: 6.0; pressure: 1 atm; temperature: 298 K ...
TRAPP was purified from 300 OD599 units of cells. SFNY904 (MATα ura3-52 bet3Δ::URA3 leu2-3, 112 BET3-protein A::LEU2 L-A-o) was used for the purification because it contained protein A-tagged Bet3p and was free of the L-A virus (L-A-o). The L-A virus coat protein, gag, is a common contaminant in the purification (see Sacher et al. 2000). To purify TRAPP, cells were converted to spheroplasts, lysed in 3 ml of buffer B (150 mM KCl, 20 mM Hepes, pH 7.2, 2 mM EDTA, 1% Trition X-100, 0.5 mM DTT, protease inhibitor cocktail), and the unbroken cells were removed as described above. The lysate was centrifuged at 14,000 g for 10 min and the supernatant (10 mg/ml of protein) was incubated with 150 μl of a 50% slurry of IgG-Sepharose (Amersham Pharmacia Biotech) for 4 h. The beads were washed three times with 3 ml of release buffer (20 mM Hepes, pH 7.2, 5 mM MgCl2, 1 mM DTT, 0.75 mM GTP, 0.75 mMGDP, 1 mg/ml BSA) or uptake buffer (20 mM Hepes, pH 7.2, 5 mM MgCl2, 1 mM EDTA, 1 mM ATP, 1 mM DTT, 1 mg/ml ...
In article ,Pine.OSF.3.95.990126164423.20371C-100000 at dingo.cc.uq.edu.au,, Paul R. Rohde ,mdprohde at mailbox.uq.edu.au, wrote: , Just a general question, , (but maybe perhaps biased towards RNA extractions). , , Whats the difference between DTT and 2ME? What does one chemical do , that the other doesnt, and when should each be used? , , How many moles of DTT equals moles of 2ME (in effectiveness)? , , Is 2ME more potent as it stinks more and more nauseating? , They are both in the lab, should know more about them. , , , , _______________________________________________________ , Paul R. Rohde http://www.uq.edu.au/~mdprohde/ * * , Department of Surgery, University of Queensland , Royal Brisbane Hospital, Queensland, 4029, Australia . * , FAX: -Australia- +61 7 3365 5559 TEL: +61 7 3362 0336 * , ______ Please reply in plain text, no html __________ Hello, I have use 2ME to break disulfide bonds in proteins (denaturing) for electrophoresis. How is it used in RNA extraction. KJ Gilbride ...
I am lysing mouse tissue samples using a dounce homogeniser in 0.5ml lysis buffer : (10%SDS, 2% Triton X100, 5M Urea, 100mM DTT and 5mM EDTA). Samples are sonicated and Gel Loading Buffer added (10% glycerol 50mM Tris pH6.8 0.2% Bromophenol blue. Samples are boiled and run on 8% SDS PAGE ...
H-Ala-cys-ser-ser-ser-pro-ser-lys-his-cys-gly-oh,(disulfide bond)/ACM888486235 can be provided in Alfa Chemistry. We are dedicated to provide our customers the best products and services.
(H-Cys-ala-oh)2,(disulfide bond)/ACM20898219 can be provided in Alfa Chemistry. We are dedicated to provide our customers the best products and services.
For sulfhydryl-selective coupling of PEG to proteins and other thiol substrates and for selective scavenging of thiol containing peptides. ...
FIG. 1. Recombinant LdmPrx shows peroxiredoxin enzyme activities in vitro. (A) Twelve percent SDS-PAGE of purified rLdmPrx under reducing conditions. Molecular mass standards are indicated on the left. (B) Western blot assay of L. donovani promastigote lysates. Cells (5 × 106) from the stationary growth phase (6 × 107/ml) were lysed directly in hot SDS sample buffer under nonreducing conditions (without DTT) and reducing conditions (in the presence of 20 mM DTT). Blots were developed with anti-LdmPrx polyclonal antibodies. Molecular mass standards are indicated on the left. (C) Nicking assay with recombinant LdmPrx was performed as described in Materials and Methods. Samples were then loaded onto a 1% agarose gel. Lane 1, only DTT; lane 2, only FeCl3; lane 3, DTT plus FeCl3; lane 4, DTT plus FeCl3 plus rLdmPrx (0.5 μM); lane 5, DTT plus FeCl3 plus rLdmPrx (1 μM); lane 6, DTT plus FeCl3 plus rLdmPrx (2 μM); lane 7, DTT plus FeCl3 plus rLdmPrx (5 μM); lane 8, DTT plus FeCl3 plus bovine serum ...
For whole cell protease treatment method, E. coli cells have been harvested, washed and resuspended in one ml Tris HCl. Proteinase K was additional to final concentrations in between 0. two mg mL 1 and 0. 5 mg mL 1 and cells had been incubated for one hour at 37 C. Digestion was stopped by washing the cells twice with Tris HCl containing 10% fetal calf serum and outer membrane proteins were ready as described over. For outer membrane proteins that have been applied for ac tivity assays, cells werent handled with Proteinase K. SDS Page Outer membrane isolates were diluted with sam ple buffer containing 4% SDS, 0. 2% bromophenol blue, 200 mM dithiothreitol and 20% glycerol boiled for 10 minutes and analyzed on 10% polyacrylamid gels. Proteins had been stained with Coomassie brilliant blue.. To correlate molecu lar masses of protein bands of curiosity, a molecular fat conventional was employed. Flow cytometer evaluation E. coli BL21 pAT kinase inhibitor MLN0128 LipBc cells were grown and ex ...
Tup1cΔ crystals were grown by hanging‐drop vapor diffusion by mixing an equal volume of protein and reservoir solution containing 50‐100 mM bis‐Tris propane pH 9, 0‐50 mM NaCl, 23‐26% (w/v) polyethylene glycol 6000 and 2 mM dithiothreitol or 1 mM BMS, and allowing the drop to equilibrate with the reservoir at 20°C. Crystals grew to an average size of 0.1 × 0.1 × 1 mm in ∼1 week. Native crystals were transferred to a drop containing well solution immediately prior to data collection. Heavy‐atom derivatives were prepared by soaking crystals in a solution containing 50 mM bis‐Tris propane pH 9 and 24% (w/v) PEG 6000 with the heavy‐atom reagent (0.5 mM EMTS for 12 h, 1 mM KAu(CN)2 for 22 h or 0.5 mM PIP for 12 h). Data were collected at room temperature on a RAXIS IIc detector equipped with a rotating‐anode Rigaku RU‐200 generator with CuKα radiation. Attempts to cryocool the crystals at −180°C were unsuccessful. All data were integrated and reduced using the ...
4-Amino-3-nitrobenzoic acid chemical properties, What are the chemical properties of 4-Amino-3-nitrobenzoic acid 1588-83-6, What are the physical properties of 4-Amino-3-nitrobenzoic acid ect.
DNA (20 pmoles) was incubated in the presence (+) or in the absence (-) of 20 pmoles of OhrR. C-Binding of OhrR. to Motif 1 buy BMS202 and Motif 2 sequences. Gel shift assay of the intergenic region and the 60 bp double strand sequences containing at their centre the genuine 17 nt corresponding to Motif 1 and Motif 2, or mutated Motif 1 with AA in place of GC (Mut1 fragment) and CCC in place of AAA (Mut2 fragment). DNA (20 pmoles) was incubated with the indicated amount of OhrR in the presence of 1 mM DTT. We took advantage of restriction sites located within the ohr-ohrR intergenic region to define further OhrR binding site. ApoI cleaved once this fragment giving a 17 bp and a 96 bp fragment. In the presence of OhrR protein the longer fragment produced two shifted bands (Figure. 3). Two HpaII sites are located within ohr-ohrR intergenic region; HpaII cleavage produced three fragments of 26, 29 and 58 bp. In the presence of OhrR, the intensity of the 58 bp fragment decreased and two retarded ...
0089]Sense and anti-sense probes were prepared from the ELOVL5 gene by incubation of the linearized gene (2 μg) with 63 μCi of [35S]UTP (1250 Ci/mmol; NEN, Massachusetts, USA) in the presence of T7 or T3 RNA polymerase. The in situ hybridization was carried out on a mouse tissue fixed with formaldehyde and embedded in paraffin. Sections (4 μm wide) were then deparaffinized in toluene and rehydrated by an alcohol gradient. After drying, the various sections were incubated in a prehybridization buffer for two hours. The hybridization was carried out overnight in a hybridization buffer (prehybridization buffer with 10 mM DTT and 2×106 cpm RNA/μl 35S-labeled) at 53° C. The excess probe was removed and the sections were inclined in an LM1 emulsion (Amersham Biosciences, UK) and exposed in the dark at 4° C. for at least one month. The sections were then developed and counterstained with hematoxylin and eosin. The hybridization with the sense probe was used as negative control and only the ...
4-Fluoro-3-nitrobenzoic acid. FC6H3(NO2)CO2H. Synonyms: . CAS 453-71-4. Molecular Weight 185.11. Browse 4-Fluoro-3-nitrobenzoic acid and related products at MilliporeSigma.
Issuu is a digital publishing platform that makes it simple to publish magazines, catalogs, newspapers, books, and more online. Easily share your publications and get them in front of Issuus millions of monthly readers. Title: DTT 1211, Author: Our City Media, Name: DTT 1211, Length: 68 pages, Page: 1, Published: 2011-11-28
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The very excellent Dr Clive Metcalfe (an ex-post-doc with me) of NIBSC has a PhD project with Prof Paul Dalby at UCL looking at disulphide bond reduction in monoclonal antibodies ...
* found in: DTT (DL-Dithiothreitol), Ultra Pure, DTT (DL-Dithiothreitol), Ultra Pure, Doxorubicin HCl, Doxycycline HCl, Reducing reagent for proteins and..
Murine N1E-115 neuroblastoma cells are shown to express a single class of angiotensin II (Ang II) receptors that display all the pharmacological properties defining the Ang II receptor subtype 2 (AT2): high affinity for 125I-labelled AT2-selective agonist CGP 42112 (Kd 91 +/- 19 pM); expected rank order of potency (CGP 42112 = (Sar1,Ile8)Ang II , or = Ang II , PD 123319 ,, DUP 753) for several Ang II analogues; increased binding in the presence of the reducing reagent dithiothreitol (DTT); and insensitivity to analogues of GTP. Molecular cloning of cDNA encoding AT2 receptors from N1E-115 cells reveals nucleotide sequence identity with the AT2 subtype expressed in fetal tissue. Murine AT2 receptors transiently expressed in COS cells display the same pharmacological profile as endogenous Ang II receptors of N1E-115 cells. Taken together, these data reveal the exclusive presence of the AT2 receptor subtype in N1E-115 cells. Incubation of N1E-115 cells with Ang II leads to a marked decrease in the ...
The abilities of cysteine-containing compounds to support growth of and influence pertussis toxin transcription, assembly, and secretion were examined. source of cysteine; however, in the absence of reduced glutathione, pertussis toxin was not efficiently secreted. Addition of the reducing agent dithiothreitol was unable to compensate for the lack of reduced glutathione and did not promote secretion of pertussis toxin. These results suggest that reduced glutathione does not affect the accumulation of assembled active pertussis toxin in the periplasm but plays a role in efficient pertussis toxin secretion by the bacterium. Pertussis toxin is a major virulence factor of heat-labile toxin, and Shiga toxin. Pertussis toxin has the most complex structure of any bacterial toxin (18, 20, 24). It is assembled from six subunits encoded by five genes, to encodes the structural gene for the A-subunit, which is an ADP-ribosyltransferase. S1 modifies mammalian G-proteins, which play a critical role in ...
We,China 2-Chloro-5-nitrobenzoic acid 2516-96-3 Suppliers and China 2-Chloro-5-nitrobenzoic acid 2516-96-3 Manufacturers, provide 2-Chloro-5-nitrobenzoic acid 2516-96-3 product and the products related with China 2-Chloro-5-nitrobenzoic acid 2516-96-3 - chinaeastchem
RhlA purification and assay.The expression of recombinant RhlA protein with an N-terminal His-tag encoded by plasmid pKZ002 was induced with isopropyl-1-thio-β-d-galactopyranoside (IPTG) in E. coli BL21(DE3). Cells were collected by centrifugation, resuspended in MCAC buffer (20 mM Tris-HCl, pH 7.9, 500 mM NaCl, 10% glycerol) and lysed with a French press. Soluble proteins were applied to a Ni2+-nitrilotriacetic acid agarose (Qiagen) column and washed with MCAC buffer plus 40 mM imidazole. His-tagged RhlA was eluted with MCAC buffer containing 200 mM imidazole. The fractions containing most of the RhlA protein were pooled, concentrated, and applied to a Superdex S200 column (GE Healthcare) to purify RhlA to homogeneity in a buffer of 20 mM Tris-HCl, pH 7.4, 1 mM dithiothreitol, 50 mM EDTA. Intact MS gave a molecular weight of 34,964, positively identifying the protein as His-tagged RhlA lacking the N-terminal fMet amino acid.. The RhlA activity was determined by measuring the formation of ...
Alfa Aesar™ 4-Acetamido-3-nitrobenzoic acid, 97+% 100g Alfa Aesar™ 4-Acetamido-3-nitrobenzoic acid, 97+% A1 to Aceto -Organics
Definition of P-hydroxymercuribenzoate with photos and pictures, translations, sample usage, and additional links for more information.
Cell isolation. PBMCs from adult and cord blood were isolated by Ficoll-Histopaque (Sigma-Aldrich) gradient centrifugation and cryopreserved in freezing medium (90% FBS + 10% DMSO; ATCC) and analyzed in batches. Fetal and adult organs were collected into cold RPMI with 10% FCS, 10 mM HEPES, penicillin, streptomycin, 0.1 mM 2-β-mercaptoethanol, 2 mM l-glutamine, and nonessential amino acids (cgRPMI medium), transported on ice, and processed within 2 hours of collection. The SI and colon were dissected from the mesentery, opened longitudinally, and cut into 1 cm sections. Mucus was removed with 3 washes in 1 mM DTT in PBS for 10 minutes. The epithelial layer was removed with 3 washes in 1 mM EDTA in PBS for 20 minutes. The intestine, MLN, liver, lung, and spleen were minced into smaller pieces and digested with freshly prepared 3 mg/mL collagenase IV (Life Technologies) and 10 mg/mL DNAse (Roche) in cgRPMI for 30 minutes, and dissociated cells were filtered through a 70 μm strainer. Cells were ...
Tubes have been incubated thirty min at 37 C during the dark. Then 1 uL of 500 mM DTT was Epothilone added to quench the alkylation response. Upcoming four. five uL of 200 mM CaCl2 were added to just about every tube. An extra five uL of 500 mM Tris HCl had been additional to sustain the pH and ionic strength. Ultimately, ten ug of trypsin or chymotrypsin dissolved in one mM HCl had been additional to every single tube. Tubes had been incubated 24 h at 37 C after which frozen at 30 C until eventually preparation for mass spectrometry. Digestion of venoms with Glu C Reduction and alkylation of venoms were carried out as described over, except that as opposed to 500 mM Tris HCl, 167 mM phosphoric acid NaOH was applied. Additionally, the enzyme was dissolved in ultrapure water, as opposed to in one mM HCl. This enabled the enzyme to cleave proteins adjacent to aspartic acid residues, likewise as glutamate residues. Once the enzyme was dissolved in one mM HCl, it cleaved subsequent to glutamate ...
Bacillus stearothermophilus 4C. SE-buffer: Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Contact: Eastsong. Phone: +86 15621028051. E-mail: [email protected] Whatsapp:+86 15621028051. Add: No.276, Zhangkou Road, Qingdao, China. ...
sample_1: unfolded apo-plastocyanin, [U-15N], 2.0 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. sample_2: unfolded apo-plastocyanin, [U-15N; U-13C], 1.5 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. sample_3: unfolded apo-plastocyanin, [U-15N; U-13C], 0.5 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. sample_4: unfolded apo-plastocyanin 0.1 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. Ex-cond_1: pH: 6.0; temperature: 308 K; ionic strength: 5 mM ...
Otavalo is a small town in Ecuador. It has about 50.000 inhabitants and is the capital of the canton of the same name. Otavalo is world-famous for its indigenous population, the so-called Otavalos, many of which are travelling around the world to sell their famous handicrafts or play in Andean Folk music groups. The Otavalos are considered the economically most successful indigenous group of Latin America, and many of the grandest houses and largest Pick-Up Trucks in Otavalo are owned by Otavalos. However, a great percentage of the Otavalos, especially in the surrounding villages, live in poverty and are victims of racial discrimination. Otavalos are easily recognized by their traditional dress: white pants and a dark poncho for men; a dark skirt and a white blouse with colourful embroidery and colourful waisteband for women. Both sexes wear their hair long (the men usually platted).
Add 1 mM fresh dithiothreitol (DTT). ACK (Ammonium-Chloride-Potassium) lysing buffer[edit]. ACK is used for lysis of red blood ...
Dithiothreitol (DTT) - used in biochemistry labs to avoid S-S bonds. *Carbon monoxide (CO) ...
In biochemistry, thiols such as β-mercaptoethanol (β-ME) or dithiothreitol (DTT) serve as reductants; the thiol reagents are ... Typically, the thiolate of a redox reagent such as glutathione or dithiothreitol attacks the disulfide bond on a protein ...
1976). "Reaction of rhodanese with dithiothreitol". Biochim. Biophys. Acta. 445 (1): 104-11. doi:10.1016/0005-2744(76)90163-7. ...
It is an epimer of dithiothreitol (DTT). The molecular formula for DTE is C4H10O2S2. Like DTT, DTE makes an excellent reducing ... Cleland, W.W. (April 1964). "Dithiothreitol, A New Protective Reagent for SH Groups". Biochemistry. 3 (4): 480-2. doi:10.1021/ ...
It also binds the sulfate ion and dithiothreitol. Maleylacetoacetate isomerase deficiency is a disease caused by a mutation in ...
biographical article Cleland, W.W. (April 1964). "Dithiothreitol, A New Protective Reagent for SH Groups". Biochemistry. 3 (4 ...
This enzyme has at least one effector, Dithiothreitol. Liau YH, Slomiany BL, Slomiany A, Piasek A, Palmer D, Rosenthal WS (1985 ...
Cleland was well known for being the first to utilize dithiothreitol for the reduction of disulfide bonds in proteins. The ... Cleland, W.W. (April 1964). "Dithiothreitol, A New Protective Reagent for SH Groups". Biochemistry. 3 (4): 480-2. doi:10.1021/ ...
It has 3 cofactors: pyridoxal phosphate, Cobamide coenzyme, and Dithiothreitol. Somack R, Costilow RN (1973). "Purification and ...
5-10 mM dithiothreitol (DTT) or β-mercaptoethanol as antioxidant. DTT is more expensive than βME, but its redox potential is ...
The thiols are then reduced with dithiothreitol and labelled by monobromobimane. Monobromobimane becomes fluorescent after ...
Another inhibitor of this enzyme is potassium pyrosulphite (K2S2O5). Banana root PPO is strongly inhibited by dithiothreitol ...
If this is not effective, dithiothreitol can be used to destroy the antibodies.:141 Cord blood samples may be contaminated with ...
Reducing conditions are usually maintained by the addition of beta-mercaptoethanol or dithiothreitol. For a general analysis of ...
Antifreeze protein Dithiothreitol, a thiol derivative of threitol Threitol at Sigma-Alrich Elks, J.; Ganellin, C. R. (1990). ...
OPAA activity is enhanced by reducing agents such as dithiothreitol (DTT) and beta-mercaptoethanol. OPAA is also catalytically ...
A23187 upregulates expression of ER stress proteins 2-deoxyglucose dithiothreitol reduces the disulfide bridges of proteins. ...
The sources of reducing power for GPx-3 in vitro include GSH, cysteine, mercaptoethanol, and dithiothreitol. There is an ...
The activity of PTGES2 is thought to be increased in the presence of sulfhydryl compounds, in particular dithiothreitol. The ...
4-dithiothreitol Thus, the two substrates of this enzyme are 2-methyl-3-phytyl-1,4-naphthoquinone and oxidized dithiothreitol, ... The systematic name of this enzyme class is 2-methyl-3-phytyl-1,4-naphthoquinone:oxidized-dithiothreitol oxidoreductase. This ... whereas its two products are 2,3-epoxy-2,3-dihydro-2-methyl-3-phytyl-1,4-naphthoquinone and 1,4-dithiothreitol. This enzyme ... oxidized dithiothreitol ⇌ {\displaystyle \rightleftharpoons } 2,3-epoxy-2,3-dihydro-2-methyl-3-phytyl-1,4-naphthoquinone + 1, ...
Dithiothreitol will slowly over time absorb more and more light in this spectrum as various redox reactions occur. Ruegg, U.T ... TCEP can keep the cysteines from forming di-sulfide bonds and unlike dithiothreitol and β-mercaptoethanol, it will not react as ... Compared to the other two most common agents used for this purpose (dithiothreitol and β-mercaptoethanol), TCEP has the ... Rhee, S. S.; Burke, D. H. (2004). "Tris(2-carboxyethyl)phosphine stabilization of RNA: comparison with dithiothreitol for use ...
However, since 2-mercaptoethanol forms adducts with free cysteines and is somewhat more toxic, dithiothreitol (DTT) is ... 2-Mercaptoethanol is often used interchangeably with dithiothreitol (DTT) or the odorless tris(2-carboxyethyl)phosphine (TCEP) ...
... effects of dithiothreitol and pepstatin A". Plant & Cell Physiology. 40 (11): 1119-26. doi:10.1093/oxfordjournals.pcp.a029496. ...
Free CoA can be regenerated from CoA disulfide and mixed CoA disulfides with reducing agents such as dithiothreitol or 2- ...
The systematic name of this enzyme class is 3-hydroxy-2-methyl-3-phytyl-2,3-dihydronaphthoquinone:oxidized-dithi othreitol ... 4-dithiothreitol. This enzyme belongs to the family of oxidoreductases, specifically those acting on the CH or CH2 groups of ... 4-dithiothreitol Thus, the two substrates of this enzyme are 3-hydroxy-2-methyl-3-phytyl-2,3-dihydronaphthoquinone and oxidized ... oxidized dithiothreitol ⇌ {\displaystyle \rightleftharpoons } 2,3-epoxy-2,3-dihydro-2-methyl-3-phytyl-1,4-naphthoquinone + 1, ...
... uses a chemical called dithiothreitol (DTT) to disrupt the sulfur bonds in the protamines in order to ...
A common 1,4-dithiol is dithiothreitol (DTT), HSCH2CH(OH)CH(OH)CH2SH, sometimes called Cleland's reagent, for to reduce protein ... a vitamin Dithiothreitol, a reagent in protein biochemistry Geminal dithiols have the formula RR'C(SH)2. They are derived from ...
This can be sidelined by either pretreatment of the erythrocytes with dithiothreitol (DTT) or by using an anti-CD38 antibody ...
Shop DL-Dithiothreitol, 99.5%, MP Biomedicals™ at Fishersci.ca ...
Dithiothreitol aka Clelands Reagent. Dithiothreitol or DTT is also known as Cleland’s Reagent. It contains two thiol groups ...
... dithiothreitol; 5azadC, 5-aza-2′-deoxycytidine; wt, wild-type HNF1 sequence; per, perfect consensus sequence; mut, mutated ...
1,4-Dithiothreitol 99.0% LR (CAS 3483-12-3), 100g. 1,4-Dithiothreitol 99.0% LR (CAS 3483-12-3), 100g.. ... 1,4-Dithiothreitol 99.0% LR (CAS 3483-12-3), 5g. 1,4-Dithiothreitol 99.0% LR (CAS 3483-12-3), 5g.. ...
The survey was a relief to feel that the addition of dithiothreitol was used for writing develop- ment programmes for other ...
DL-Dithiothreitol [DTT], Cat#A2452 Quick view Wishlist DL-Dithiothreitol [DTT], Cat#A2452 ...
DL-Dithiothreitol [DTT], Cat#A2452 Quick view Wishlist DL-Dithiothreitol [DTT], Cat#A2452 ...
Dive into the research topics of An optimized protocol for isolation of soluble proteins from microalgae for two-dimensional gel electrophoresis analysis. Together they form a unique fingerprint. ...
Dithiothreitol-based oxidative potential for airborne particulate matter: an estimation of the associated uncertainty. Molina, ... Among them, the dithiothreitol (DTT) assay has gained popularity due to its simplicity and overall low implementation cost. U ...
DTT (Dithiothreitol) [3483-12-3] Availablility: In Stock View Product ID: 19733320 ...
Dithiothreitol (Sigma Aldrich; ≥99.0%; 43819). Equipment. *Sampling tubes resistant to snap-freezing in liquid nitrogen (Falcon ...
50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol ...
50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol ...
50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol ...
1 mM dithiothreitol, 4 U/ml RNAse inhibitor cocktail and protease inhibitors (Roche, used at 1× concentration, as specified by ...
DL-dithiothreitol (DTT). Psychopharmacology 174, 283-290. Dalvi, A. and Rodgers, R. J. (1996) GABAergic influences on plusmaze ...
Dithiothreitol, and Glycerol. ...
Our results indicate that addition of dithiothreitol improves glutenin extraction without interfering with protein secondary ...
The method is based on the catalytic effect of selenium on the reduction of tetranitro blue tetrazolium by dithiothreitol; it ... N2 - The method is based on the catalytic effect of selenium on the reduction of tetranitro blue tetrazolium by dithiothreitol ... AB - The method is based on the catalytic effect of selenium on the reduction of tetranitro blue tetrazolium by dithiothreitol ... "The method is based on the catalytic effect of selenium on the reduction of tetranitro blue tetrazolium by dithiothreitol; it ...
5. 無臭 - 無惡臭氣味,且不添加2-mercaptoethanol (2-ME)或Dithiothreitol (DTT)。. 二、產品資訊. 貨號. 品名.
... the efficient CPT launch habits of CPT-LA CNM was confirmed within the decreasing circumstances containing dithiothreitol (DTT ...
Transforming growth factor-beta induces formation of a dithiothreitol-resistant type I type II receptor complex in live cells ...
After that, we reduce the sample with 1 M dithiothreitol (DTT) for 30 minutes, and add Alkylate in 0.5 M â ¦ 1 . USP Labs ...
... and a drastic redistribution between the mass concentration and OP measured by both ascorbic acid and dithiothreitol is ...
Unit calculation assay conditions: 100 mM NaCl, 50 mM Tris-HCl (pH 7.9 @ 25°C), 10 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml BSA ... Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA and 50% glycerol. Store at ...
Unit calculation assay conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9 @ 25oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml ... Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml bovine serum albumin and 50% ...
Exogenously added thiols L-cysteine or dithiothreitol inhibited the relaxant responses to FPTO but not to sodium nitroprusside ...
... facilitates the preparation of PDMA-based polymersomes capable of dithiothreitol-induced pyrene release as evidenced by ... of dithiothreitol as determined by UV−vis spectroscopy. ...
10 mM fresh dithiothreitol, 5 mM of each ribonucleotide triphosphate, 100 g/ml T7 Pol and 1 M DNA template. The response was ...
  • Dithiothreitol (DTT) is the common name for a small-molecule redox reagent also known as Cleland's reagent. (wikipedia.org)
  • Dithiothreitol, DTT or Cleland's reagent is a small-molecule redox reagent. (glentham.com)
  • Dithiothreitol shelf life can be extended with refrigeration at 2-8 °C. Oxidation presents further complications as oxidized DTT exhibits a strong absorbance peak at 280 nm. (wikipedia.org)
  • Due to air oxidation, DL-dithiothreitol (DTT) is a relatively unstable compound whose useful life can be extended by refrigeration and handling in an inert atmosphere. (24marketreports.com)
  • Dithiothreitol (DTT) is the common name of the more popular of the two Cleland's Reagents (the other being Dithioerythritol or DTE). (goldbio.com)
  • EDTA ), and reducing agents like dithiothreitol (DTT). (wikipedia.org)
  • N-acetylcysteine (NAC) and Dithiothreitol (DTT), two compounds that prevent experimental amyotrophic lateral sclerosis (ALS) in the vitamin C - deficient guinea pig[1], were given to patients with motor neuron disease (MND). (springer.com)
  • The reduction of disulfide-containing compounds by Dithiothreitol is a two-step reaction. (cfmot.de)
  • Dithiol compounds such as dithiothreitol inhibited AA release from both the thimerosal- and the PAO-treated cells, and monothiol compounds (l-Cys and glutathione) decreased the thimerosal response. (biomedsearch.com)
  • Dithiothreitol markedly increased the ligand binding affinity of angiotensin II (AII) receptor type II (AT 2 ) without affecting its antagonist selectivity in cultured ovarian granulosa cells, demonstrating that this AT 2 is of the dithiothreitol-sensitive type. (elsevier.com)
  • Most commonly, Dithiothreitol is used to reduce disulfide bonds in peptides and proteins. (cfmot.de)
  • The most commonly used reductant is dithiothreitol (DTT). (thomassci.com)
  • Dithiothreitol" is a descriptor in the National Library of Medicine's controlled vocabulary thesaurus, MeSH (Medical Subject Headings) . (harvard.edu)
  • Dithiothreitol is an integral reagent, together with denaturants such as GdmCl, for the solubilization of proteins from inclusion bodies. (cfmot.de)
  • In consumption market, the global consumption value of DL-dithiothreitol (DTT) increases with the 4.55% average growth rate. (24marketreports.com)
  • To study and analyze the global DL-Dithiothreitol (DTT) consumption (value & volume) by key regions /countries, product type and application, history data from 2014 to 2018, and forecast to 2024. (24marketreports.com)
  • The protective effect of hypoxia and dithiothreitol on X-ray-induced genetic damage in Arabidopsis. (wur.nl)
  • Dithiothreitol repairs up to 30% of DNA radicals, but an increase in the thiol concentration above 10 mM, even up to 1000 mM, does not further enhance the overall level of the repair process. (warwick.ac.uk)
  • To understand the structure of DL-Dithiothreitol (DTT) market by identifying its various subsegments. (24marketreports.com)
  • Effect of zinc and dithiothreitol (DTT) on LLER oligomerization. (nih.gov)
  • The effect of a thiol, dithiothreitol (DTT), on DNA radiolysis at cryogenic temperatures was investigated by the EPR method. (warwick.ac.uk)
  • Dithiothreitol is the trans isomer of the compound 2,3-dihydroxy-1,4-dithiolbutane. (cfmot.de)
  • High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol. (nih.gov)
  • We conclude that NAT1 expression is not directly regulated by E 2 , DHT, 3 β -adiol, or dithiothreitol despite high NAT1 and ESR1 expression in luminal A breast cancer cells, suggesting that ESR1 , XBP1, and NAT1 expression may share a common transcriptional network arising from the luminal epithelium associated with better survival in breast cancer. (aspetjournals.org)
  • According to this study, over the next five years the DL-Dithiothreitol (DTT) market will register a 2.5% CAGR in terms of revenue, the global market size will reach US$ 1080 million by 2024, from US$ 950 million in 2019. (24marketreports.com)
  • With reducing action of DL-dithiothreitol (DTT), the downstream application industries will need more DL-dithiothreitol (DTT) products. (24marketreports.com)
  • Herein, we used low-dose pharmacological UPR inducers such as tunicamycin (TM) and dithiothreitol (DTT) to efficiently activate the IRE1-XBP-1 pathway in myeloma cells characterized by transcriptional expression increase in spliced XBP-1 and molecular chaperons, accompanied by significant differentiation and maturation of these myeloma cells, without concomitant cytotoxicity. (sigmaaldrich.com)
  • Dithiothreitol (DTT) effectively suppressed the phenylarsine oxide-inhibited cellular reductive capacity, but unexpectedly, enhanced As 2 O 3 -induced apoptosis in NB4 cells. (aspetjournals.org)
  • Dithiothreitol increased levels of the activated, spliced XBP1 in ER α (+) MCF-7 and T47D breast cancer cells but did not affect NAT1 or ESR1 expression. (aspetjournals.org)
  • This graph shows the total number of publications written about "Dithiothreitol" by people in Harvard Catalyst Profiles by year, and whether "Dithiothreitol" was a major or minor topic of these publication. (harvard.edu)
  • International validation of a dithiothreitol (DTT)-based method to resolve the daratumumab interference with blood compatibility testing. (harvard.edu)
  • DL-Dithiothreitol Market also splits the market by region: Breakdown data in Chapter 4, 5, 6, 7 and 8. (24marketreports.com)
  • The key manufacturers covered in DL-Dithiothreitol Market : Breakdown data in in Chapter 3. (24marketreports.com)
  • In addition, DL-Dithiothreitol Market discusses the key drivers influencing market growth, opportunities, the challenges and the risks faced by key manufacturers and the market as a whole. (24marketreports.com)
  • Acid treatment of influenza A and B virus preparations followed by addition of dithiothreitol (DTT) and centrifugation through a sucrose cushion removes the HA1 subunit of hemagglutinin from virus. (nih.gov)
  • DL-Dithiothreitol Market presents a comprehensive overview, market shares, and growth opportunities of DL-Dithiothreitol (DTT) market by product type, application, key manufacturers and key regions and countries. (24marketreports.com)
  • Dithiothreitol is useful for specifically detecting low levels of the AT 2 in the ovary, where it plays roles that are probably related to atresia. (elsevier.com)
  • Chisholm KM, Getsinger D, Vaughan W, Hwang PH, Banaei N. Pretreatment of sinus aspirates with dithiothreitol improves yield of fungal cultures in patients with chronic sinusitis. (harvard.edu)