Such mixtures of amphoteric electrolytes or buffers that provide a continuous range of pH in an electric field; used for separating proteins by their isoelectric points, i.e., by isoelectric focusing.
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.

Adenosine-5'-O-phosphorylated and adenosine-5'-O-phosphorothioylated polyols as strong inhibitors of (symmetrical) and (asymmetrical) dinucleoside tetraphosphatases. (1/14)

Dinucleoside 5',5"'- P (1), P ( n )-polyphosphates, and particularly the diadenosine compounds, have been implicated in extracellular purinergic signalling and in various intracellular processes, including DNA metabolism, tumour suppression and stress responses. If permitted to accumulate, they may also be toxic. One approach to understanding their function is through the various specific degradative enzymes that regulate their levels. Eight adenosine-5'- O -phosphorylated polyols (derivatives of glycerol, erythritol and pentaerythritol) and 11 adenosine-5'- O -phosphorothioylated polyols (derivatives of glycerol, erythritol, pentaerythritol, butanediol and pentanediol) have been tested as inhibitors of specific diadenosine tetraphosphate (Ap(4)A) hydrolases. Of these two groups of novel nucleotides, the adenosine-5'- O -phosphorothioylated polyols were generally stronger inhibitors than their adenosine-5'- O -phosphorylated counterparts. 1,4-Di(adenosine-5'- O -phosphorothio) erythritol appeared to be the strongest inhibitor of ( asymmetrical ) Ap(4)A hydrolases (EC 3.6.1.17) from both lupin and human, with K (i) values of 0.15 microM and 1.5 microM respectively. Of eight adenosine-5'- O -phosphorylated polyols, 1,4-di(adenosine-5'- O -phospho) erythritol was the only compound that inhibited the lupin enzyme. Two derivatives of pentaerythritol, di(adenosine-5'- O -phosphorothio)-di(phosphorothio) pentaerythritol and tri(adenosine-5'- O -phosphorothio)-phosphorothio-pentaerythritol, proved to be the strongest inhibitors of the prokaryotic ( symmetrical ) Ap(4)A hydrolase (EC 3.6.1.41) so far reported. The estimated K (i) values were 0.04 microM and 0.08 microM respectively. All of these inhibitors were competitive with respect to Ap(4)A. These new selectively acting Ap(4)A analogues should prove to be valuable tools for further studies of Ap(4)A function and of the enzymes involved in its metabolism.  (+info)

Efficient solubilization buffers for two-dimensional gel electrophoresis of acidic and basic proteins extracted from wheat seeds. (2/14)

Plant tissues are made up of a broad range of proteins with a variety of properties. After extraction, solubilization of a diverse range of plant proteins for efficient proteomic analysis using two-dimensional electrophoresis is a challenging process. We tested the efficiency of 12 solubilization buffers in dissolving acidic and basic proteins extracted from mature seeds of wheat. The buffer containing two chaotropes (urea and thiourea), two detergents (3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propane-sulfonate and N-decyl-N,N-dimethyl-3-ammonio-1-propane-sulfonate), two reducing agents (dithiothreitol and tris (2-carboxyethyl) phosphine hydrochloride) and two types of carrier ampholytes (BioLyte pH 4-6 and pH 3-10) solubilized the most acidic proteins in the pH range between 4 and 7. The buffer made up of urea, thiourea, 3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propane-sulfonate, DeStreak reagent (Amersham Biosciences, Uppsala, Sweden) and immobilized pH gradient buffer, pH 6-11 (Amersham Biosciences) solubilized the most basic proteins in the pH range between 6 and 11. These two buffers produced two-dimensional gels with high resolution, superior quality and maximum number of detectable protein (1425 acidic protein and 897 basic protein) spots.  (+info)

Commercial ampholytes used for isoelectric focusing may interfere with bioactivity based purification of antimicrobial peptides. (3/14)

BioRad's Rotofor system has been frequently used for the purification of proteins and smaller peptides such as bacteriocins. In this study, we report that some commercially available ampholytes used with the Rotofor isoelectric focusing system possess antimicrobial activity, which may interfere with the purification of bacteriocins and bacteriocin-like substances.  (+info)

Infection with Panton-Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus t034. (4/14)

 (+info)

Effect of synthetic carrier ampholytes on saturation of human serum transferrin. (5/14)

We have investigated the effect in solution of synthetic carrier ampholytes on the saturation of human serum transferrin. By spectrophotometric titrations of human serum transferrin with various Fe3+-carrier ampholyte solutions, we demonstrated that under these conditions carrier ampholytes behave as typical chelators, their binding curves being very similar to that obtained with disodium nitrilotriacetate. On performing titration experiments at three different pH values, carrier ampholytes act like nitrilotriacetate at pH 7.5, but the former are more effective iron donors at pH 8.4 and worse iron donors at pH 5.2. Spectrophotometric titrations of isolated C-terminal and N-terminal fragments obtained from human serum transferrin by thermolysin cleavage show no differences between them, and no differences with respect to the whole protein except that they contain half the number of binding sites. In order to determine a site-specificity of iron in the presence of ampholytes, the classical urea/polyacrylamide-gel-electrophoresis technique was adopted. Under saturating conditions carrier ampholyte solutions act mostly on the C-terminal site, whereas desaturating agents remove iron preferentially from the N-terminal site. Our findings support the hypothesis that Ampholine may chelate Fe3+ as well as many other compounds.  (+info)

Resolution of alkaline phosphatase isoenzymes in serum by isoelectric focusing in immobilized pH gradients. (6/14)

This new method for fractionation of serum alkaline phosphatase isoenzymes is based on isoelectric focusing on a mixed-type polyacrylamide support containing an immobilized pH gradient with a superimposed carrier-ampholyte gradient. All known forms of alkaline phosphatase are separated in an Immobiline pH 3.5-6.0 gradient, the sample being applied into pockets cast on a pH 8.0 plateau. Sharp zymogram bands are obtained by substituting alkaline-stable 5-bromo-4-chloro-3-indoxyl phosphate and tetrazolium salts for the standard 1- and 2-naphthyl phosphate-diazonium salt combinations. After hydrolysis of the phosphate group by the alkaline phosphatase the indoxyl moieties reduce tetrazolium salts to nearly insoluble and nondiffusible formazan precipitates. Normal sera show an array of about 10 isobands isoelectric between pH 3.9 and pH 4.79. In Paget's disease, two sharp isobands with pls of 4.97 and 5.09 are seen. Placental alkaline phosphatase overlaps with the higher pl bands of normal serum; however, upon heat destruction of the latter, it shows four sharp bands with the following pl's: 4.59, 4.62, 4.67 and 4.73.  (+info)

Charge microheterogeneity of the major capsid protein of polyoma virus. (7/14)

The behavior in isoelectric focusing of the major capsid polypeptide VPI of several strains of polyoma virus was studied. Two previously recognized phenomena were reexamined, namely, (i) the separation of the VP1 polypeptide into multiple subspecies differing only slightly from each other in apparent isoelectric point and (ii) strain differences in the overall apparent net charge of the family of VP1 subspecies. It was found that the pattern of subspecies was reproducible when focusing was initiated from either the basic or acidic region of the gel, keeping the ampholyte mixture constant. However, individual subspecies were unstable, and labeled polypeptide could be shifted dramatically by either refocusing of separated subspecies or by altering the concentration of ampholytes. These findings suggest that protein-protein and protein-ampholyte interactions play an important role in the generation of this charge heterogeneity. The basis for the overall charge difference between the VP1 of 3049 virus and several other strains (lpD, lpS, ts59, and A2) was studied, using recombinant viruses constructed of specific sequences derived from 3049 and lpD genomes. The portion of the VP1 polypeptide carrying the altered charge could be mapped to the body of the molecule 3' to the HindIII site at 45.0 map units (3,918 base pairs). This clearly segregates the VP1 charge phenotype from the cyc phenotype of 3049 in which capsid proteins are overproduced and accumulate in the cytoplasm of infected cells.  (+info)

pH-dependent aggregation and electrofocusing of poliovirus. (8/14)

Following isoelectric focusing, poliovirus can be detected at two pH values. The acidic form consists of poliovirus aggregates and the neutral form of single virions.  (+info)

Ampholyte mixtures are a type of chemical compound that contain both acidic and basic groups within the same molecule. They are commonly used in a process called isoelectric focusing, which is a technique used to separate and purify proteins and other large molecules based on their isoelectric points (pI).

An ampholyte mixture typically contains a range of amphoteric compounds with different pI values, which create a pH gradient when subjected to an electric field. This gradient allows proteins and other large molecules to migrate to a position in the gradient where their net charge is zero (their isoelectric point), effectively separating them based on their pI.

Ampholyte mixtures are usually composed of a complex mixture of organic compounds, including polyamino polycarboxylic acids and their salts. They are available in various ranges of pH values to suit different applications.

Isoelectric focusing (IEF) is a technique used in electrophoresis, which is a method for separating proteins or other molecules based on their electrical charges. In IEF, a mixture of ampholytes (molecules that can carry both positive and negative charges) is used to create a pH gradient within a gel matrix. When an electric field is applied, the proteins or molecules migrate through the gel until they reach the point in the gradient where their net charge is zero, known as their isoelectric point (pI). At this point, they focus into a sharp band and stop moving, resulting in a highly resolved separation of the different components based on their pI. This technique is widely used in protein research for applications such as protein identification, characterization, and purification.

Neutral Form Equilibrium of Ampholytes in Pure Solvents and Mixtures". J. Am. Chem. Soc. 119 (21): 4999-5006. doi:10.1021/ ... they sometimes have different enough structures that they can be detected independently in their mixture. This allows ...
... is a protein-separation technique that allows resolution of single proteins and other ampholytes from a complex mixture ...
... ampholyte mixtures MeSH D27.720.470.305 - culture media MeSH D27.720.470.305.250 - culture media, conditioned MeSH D27.720. ...
Immediate precipitation may occur giving a cloudy mixture. The solubility measured for such a mixture is known as "kinetic ... Solubility values of organic acids, bases, and ampholytes of pharmaceutical interest may be obtained by a process called " ... In static methods a mixture is brought to equilibrium and the concentration of a species in the solution phase is determined by ...
... s (IPG) are made by mixing two kinds of acrylamide mixture, one with Immobiline having acidic buffering ... method eliminates the problems of gradient instability and poor sample loading capacity associated with carrier ampholyte pH ...
When a sample (a mixture of peptides or proteins) is injected in the capillary, the presence of the electrical field and the pH ... Molecules to be focused are distributed over a medium that has a pH gradient (usually created by aliphatic ampholytes). An ... IEF involves adding an ampholyte solution into immobilized pH gradient (IPG) gels. IPGs are the acrylamide gel matrix co- ... The multi-junction IEF system has been used to separate tryptic peptide mixtures for two-dimensional proteomics and blood ...
This method only gives a rough measure of the amounts of different proteins in the mixture, and it is not able to distinguish ... By making use of a pH-gradient, that can for example be induced by ampholytes, this technique allows to separate protein ... Protein purification is a series of processes intended to isolate one or a few proteins from a complex mixture, usually cells, ... When a vessel (typically a tube or bottle) containing a mixture of proteins or other particulate matter, such as bacterial ...
ISBN 978-1-4200-7099-6. Pagliara A, Carrupt PA, Caron G, Gaillard P, Testa B (1997). "Lipophilicity Profiles of Ampholytes". ... is the ratio of concentrations of a compound in a mixture of two immiscible solvents at equilibrium. This ratio is therefore a ...
Neutral Form Equilibrium of Ampholytes in Pure Solvents and Mixtures". J. Am. Chem. Soc. 119 (21): 4999-5006. doi:10.1021/ ... they sometimes have different enough structures that they can be detected independently in their mixture. This allows ...
5. Other components include dyes, combs, gel mixtures, gel dryers and chemicals such as denaturing agents, gel hardeners, and ... ampholytes. *6. . Conventional electrophoresis *7. . Principle of Electrophoresis • In the electric field the negative end ...
Carrier ampholytes help to stabilize the pH gradient and current in IPG strips and aid in protein solubility, resulting in more ... The buffer typically contains a denaturing agent, urea or mixture of urea and thiourea, a non-ionic or zwitterionic detergent, ... Carrier ampholytes help to stabilize the pH gradient and current in IPG strips and aid in protein solubility, resulting in more ... The ZOOM Carrier Ampholytes are small, soluble molecules with both positive and negative charge groups that sort at their ...
The ampholyte mixture was adapted (i.e. nature and concentration of carrier ampholytes, volume of sacrificial electrolytes) in ... Catechol was specifically preconcentrated in a pH 8.5 phosphate buffer/MeOH (80/20, v/v) mixture. A volume up to 20 times the ... We report that the LC-MS-IRMPD approach offers a robust advanced MS diagnostic of mixtures of isomers, including carbohydrate ... Fractionation of the CS disaccharides mixture in isolated ITP bands and collection of solutes were successfully done using a ...
... and Bio-Lyte 3-10 Ampholyte (0.2%) (Bio-Rad, Cat. No.163-1113). The protein mixture was then applied to ReadyStrip™ IPG 7 cm ...
Ampholyte Mixtures. Amphotericin B. Ampicillin. Ampicillin Resistance. Amplifiers. Amprolium. Ampulla of Vater. Amputation. ...
... ampholytes (40% BioLyte® 3/10 Ampholyte, Bio-Rad Laboratories, Inc., Hercules, CA, USA) to adjust to a final volume of 330 μL ... The third and the fourth groups were mixtures of Labofeed H feed with the addition of vegetable oils with high PUFAs content, ... Real-time PCRs were conducted in the 20 μL mixture contained 5 μL of diluted cDNA (dilution factor 25), 200 nM each of the ... who reported that fish oil mixture supplementation of a high-fat high-sucrose (HFHS) diet significantly downregulated HGD level ...
educational interviews, and High mixtures. required dextran, a Domestic Sketch, In One agent By Hattie L. Domestic Fecility, a ... Basamak 2004 were that this impact does less unterdrü to slavery from changes and ampholytes than additional separate studies( ... phospholipids are exclude our phase ampholytes. CloseLog InLog In; ebook Beggars of Life: A Hobo; FacebookLog In; vereinbart; ...
Mixtures with other solvents. Ethanol is miscible with water in any ratio. In this case, volume contraction occurs during ... Ethanol can react as both a Brønsted acid and a Brønsted base, making it an ampholyte: 2 C2H5OH → C2H5OH2+ + C2H5O-. The ... This three-substance mixture of ethanol, water and entrainer enters the auxiliary separation column. There it is separated into ... The total volume of a water/ethanol mixture is smaller than the sum of the individual volumes. Thus, mixing 50 mL of ethanol ...
Modification of Polyethylene by RF Plasma in Different/Mixture Gases. E. Abusrafa A, Habib S, Krupa I, Ouederni M, Popelka A. ... Preparation of an Interpenetrating Network of a Poly(ampholyte) and a Cholesteric Polymer and Investigation of Its Hydrochromic ...
The cells were incubated at 37°C in a humidified gas mixture containing 5% CO2 balanced with air. Chondrocytes were used at ... carrier ampholytes (IPG Buffer pH 3-10 NL), 0.4% Triton X-100 and 2 mM dithiothreitol (DTT). ...
4 µl of the Cy2 solution is added to the internal standard protein mixture, which contains 12.5 µg protein from each sample. ... ampholyte 1% * Volume of loading buffer: 35 µL Loading method: cup loading. 3.3 Protocol 3.3.1 Buffers No buffer. 3.3.2 ... ampholyte 1% 3. Gel matrix and electrophoresis protocol ------------------------------------------ 3.1 Dimension details 3.1.1 ...
A solution of this issue is based on data on the vapor-liquid and liquid-liquid equilibria of the mixture and its components, ... article shows the principal possibility of identification of EPO-Fc fusion protein by means of IEF-PAGE in carrier ampholyte- ... A strategy of studying the three-liquid phase equilibrium area in quaternary mixtures is proposed. The strategy is based on the ... It is shown that the Ta-Re/Al2O3 catalyst allows transforming ethanol and mixtures of ethanol with glycerin into aliphatic ...
To prepare 100 ml of a 6 % acrylamide gel, 20 ml of the 30 % acrylamide stock solution is added into the gel-mixture.. Of ... but only after Vesterberg successfully synthesized carrier ampholytes for generation of a continuous pH gradient, the technique ... The lower the ratio of these two components in the mixture, the higher the degree of crosslinking. This means that a 6 % gel ... In contrast to the todays methods, a complete separation of the components of a mixture was never achieved, no matter how long ...
两性电解质, Pharmalyte, Ampholyte*琼脂糖凝胶, Cytiva Sepharose*葡聚糖凝胶, Cytiva Sephadex*超敏葡琼凝胶, Cytiva Superdex*标签蛋白亲和纯化介质, Tag-Medium*抗体纯化 ... To compose a solvent mixture with diethyl ether for the purification
Westall, R.G. 1952: The amino-acids and other ampholytes of urine. 1. A general method of isolation. Biochem. J 52: 638-642 ... A general technique applied to the estimation of cyanocobalamin and hydroxycobalamin in mixtures of the vitamins. Chem and ... Westall, R.G. 1955: The amino acids and other ampholytes of urine. 3. Unidentified substances excreted in normal human urine. ... Lysenko, T.D. 1954: The application of mixtures of organic and mineral fertilizers under winter cereals. Agrobiologiya 4, 7-13 ...
Ampholyte Mixtures Actions. * Search in PubMed * Search in MeSH * Add to Search ...
Fetal exposure to chlordane and permethrin mixtures in relation to inflammatory cytokines and birth outcomes.Environ Sci ... was performed without ampholytes and glycerol according to the manufacturers instructions for peptide focusing. In short, the ... and the mixture was incubated at 37°C overnight. Tryptic digests (100-500 μg) were then fractionated with the Agilent 3100 ...
Ampholyte Ampholyte Mixture Ampholytes Carrier Ampholyte Carrier Ampholytes Isoelectric Focusing Agent Isoelectric Focusing ... Ampholyte Mixtures Preferred Concept UI. M0001020. Registry Number. 0. Related Numbers. 70322-21-3. Scope Note. Such mixtures ... Ampholytes Narrower Concept UI. M0003008. Registry Number. 0. Terms. Ampholytes Preferred Term Term UI T005735. Date09/18/1980 ... Ampholyte Mixtures Preferred Term Term UI T002009. Date01/01/1999. LexicalTag NON. ThesaurusID NLM (1980). ...
Ampholyte Ampholyte Mixture Ampholytes Carrier Ampholyte Carrier Ampholytes Isoelectric Focusing Agent Isoelectric Focusing ... Ampholyte Mixtures Preferred Concept UI. M0001020. Registry Number. 0. Related Numbers. 70322-21-3. Scope Note. Such mixtures ... Ampholytes Narrower Concept UI. M0003008. Registry Number. 0. Terms. Ampholytes Preferred Term Term UI T005735. Date09/18/1980 ... Ampholyte Mixtures Preferred Term Term UI T002009. Date01/01/1999. LexicalTag NON. ThesaurusID NLM (1980). ...
Ampholyte. Ampholyte Mixture. Ampholyte, Carrier. Ampholytes. Ampholytes, Carrier. Carrier Ampholyte. Carrier Ampholytes. ... Ampholyte Mixture Ampholyte, Carrier Ampholytes, Carrier Carrier Ampholyte Carrier Ampholytes Focusing Agent, Isoelectric ... Ampholyte Mixtures - Preferred Concept UI. M0001020. Scope note. Such mixtures of amphoteric electrolytes or buffers that ... Ampholyte Mixtures Entry term(s). Agent, Isoelectric Focusing Agents, Isoelectric Focusing Ampholines ...
In this procedure, a red blood cell hemolysate is focused on a 5% agarose gel containing ampholytes of a 5 to 7 pH range. The ... In isoelectric focusing, a pH gradient is established across an agarose gel by adding a select mixture of amphoteric molecules ... isoenzyme bands are then visualized by applying a substrate mixture that results in a series of reactions (shown below). The ...
BRONCHODILATOR AGENTS AMPHOLYTE MIXTURES BUFFERS BICARBONATES BUFFERS BUFFERS BUFFERS HEPES BUFFERS TROMETHAMINE BUFFERS BAY-K- ...
In mixtures of the two acetic fixation (simultaneously)is evident, but some nucleoli in some nuclei stain. In some instances ... there probably is some ampholyte present un-doubtedly a protein or proteins. This protein is not nucleoprotein, as it is ... The author, at another laboratory, has investigated, to some extent, the effects of fixatives, separately and in mixtures, on ... in a mixture.These data concern the penetration of fixatives [?] into pieces of a guinea pig liver. This authors experience ...
Ampholytes. Amphoteric electrolytes, the most common of which are amino acids, peptides, and proteins, can react as both acids ... a concentrated mixture of polycyclic pancuronium+ and aromatic pentothal- opposite ions). ... Ampholytes are least soluble at their isoelectric pH, pI, or zwitterion point, which equals the mean of the pKa values for ...
Figure 9.3.8) and do not stain nucleic acids, which are sometimes found in protein mixtures from cell or tissue extracts.. In ... such as ampholyte staining or mirroring effects on the plastic gel backing. ... Figure 9.3.8 Quantitation of proteins in a gel using SYPRO Orange protein gel stain (S6650, S6651). A protein mixture was ... We offer a protein mixture for use as molecular weight markers in SDS-polyacrylamide gel electrophoresis (Figure 9.3.12). This ...
In relatively going, they are a download Использование Adobe Dreamweaver CS4 of normal mixtures altered along the time of the ... A download of certain ampholytes recovered adjusted for using cookies. thereby, more protonated or more unbound logistics of ... Composition screenplay has for the shared performance of graduate ampholytes of performance onto the separating fool, compared ... the degree of the first laboratories and the mixture Fig. is important. The Book of trials to be isolated is followed as a ...
Mixtures of PEG and a large model protein, glucose isomerase, have been studied to characterize the effect of a metastable ... A novel ampholyte buffer suitable for crystallization using the ProTeam FFE apparatus has been developed. This buffer is able ... Development of a novel ampholyte buffer for isoelectric focusing: electric charge-separation of protein samples for X-ray ...
Sinus and congestion mixture of three drugs and an impurity was studied for their spectral resolution using four multivariate ... Most of them are ampholytes with one chiral center. Stereogeneic center is located either in the side ring of Gatifloxacin ( ... Concentration levels in the designed mixture were carefully considered to recede the challenging dosage form ratio. The value ... Spectral resolution of quaternary components in a sinus and congestion mixture; Multivariate algorithms to approach extremes of ...
Bio-Rad伯樂兩性電解質 Bio-Lyte 3/10 Ampholyte 40%, 25 ml (1631113) ... Wako表面濕潤張力測試混合溶液 Wetting Tension Test Mixture ... Wako表面濕潤張力測試混合溶液 Wetting Tension Test Mixture ... WAKO
Considering a general case (based on the derivations for mono- and diprotic acids and bases and ampholytes) this formula can be ... Unordered Mixtures and Ordered Mixtures. *Charge of the Group. *Markush Structures *R-groups ...
  • The buffer typically contains a denaturing agent, urea or mixture of urea and thiourea, a non-ionic or zwitterionic detergent, a reducing agent to cleave disulfide-bonds, and ampholytes. (thermofisher.com)
  • Urea 7M Thiourea 2M Chaps 4% ampholyte 1% 3. (expasy.org)
  • Two-dimensional gel electrophoresis (2DE) is an established technique for high-resolution profiling of complex protein mixtures. (thermofisher.com)
  • 4 µl of the Cy2 solution is added to the internal standard protein mixture, which contains 12.5 µg protein from each sample. (expasy.org)
  • The lower the ratio of these two components in the mixture, the higher the degree of crosslinking. (itwreagents.com)