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Dominic-Luc Webb molmed wrote: , On Thu, 20 Dec 2001, Duncan Clark wrote: , , , ,In most of the places varying concentration of IPTG has been suggested , , ,for induction depending upon the expression system and other things. , , ,Most of these are for an low ODs like 0.5-1. , , , , , ,In case we need to work on high ODs like 50-100 or 150 would one , , , You will never get that far for the simple reason that , spectrophotometers peak out at about 3. They will give , values up to about 4, which are meaninglessly beyond , the linear range of the spectrophotometer. OD600 are , absorbance values. Yes and no. I also think bugs cannot grow at so high OD600 (at least E. coli or similar gram negative bacteria). However, its very common to get OD600 near 6 or 7, depending on the media. To measure it, the sample must be diluted in fresh media to avoid the saturation of the spectrophotometer. Usually, we dilute the culture to get a OD 600nm between 0.1 and 1.5 Your bugs will already be dying , at about ...
Author(s): Ruegg, Thomas L; Pereira, Jose H; Chen, Joseph C; DeGiovanni, Andy; Novichkov, Pavel; Mutalik, Vivek K; Tomaleri, Giovani P; Singer, Steven W; Hillson, Nathan J; Simmons, Blake A; Adams, Paul D; Thelen, Michael P | Abstract: In the original version of this Article, an incorrect URL was provided in the Data Availability Statement regarding the deposition of plasmids listed in Supplementary Table 4. The correct URL is https://public-registry.jbei.org/folders/378 . This error has been corrected in both the PDF and HTML versions of the Article.
im not sure what you mean exactly when you say question in cell disruption and making sure youre protein is as soluble as possible. usually you need to play around with IPTG concentration and try to lower the induction temp a little to try and maximise the yeild of soluble protein. i am assuming you are doing iptg induction as there arent any details ...
This page contains information on the chemical 1,3-Propanediol, 2-(hydroxymethyl)-2-isopropyl-1-methyl-, cyclic phosphate (1:1) including: 5 synonyms/identifiers.
1-(tert-butyl)-2-isopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-imidazole; CAS Number: 2223040-48-8; find CombiPhos Catalysts Inc-CO1H324A64E0 MSDS, related peer-reviewed papers, technical documents, similar products & more at Sigma-Aldrich
You are viewing an interactive 3D depiction of the molecule 1-isopropyl-6-methylergoline-8-carboxylic acid (C19H24N2O2) from the PQR.
6,7-dihydro-4-hydroxy-7-isopropyl-6-oxo-N-(3-(piperidin-1-yl)propyl)thieno(2,3-b)pyridine-5-carboxamide: VRX-03011 is the potassium salt; structure in first source
Structure, properties, spectra, suppliers and links for: N-[(10S,13S,20R)-3,35-Dichloro-18,21-dihydroxy-10-isopropyl-12-oxo-8,22,39-trioxa-4,11,34,38-tet.
Structure, properties, spectra, suppliers and links for: (3S,5R,6E)-7-[3-(4-Fluorophenyl)-1-isopropyl-1H-indol-2-yl]-3,5-dihydroxy-6-heptenoate.
To determine whether the mutation of Tyr515 could affect to the activity of AtSSIV, we cloned the two mutated versions into the pDEST17 vector, which allow the IPTG-inducible expression of the SSIV versions (AtSSIV-Y515F-WCTP, AtSSIV-Y515E-WCTP). In addition, this vector allows the tagging of the polypeptides expressed with a His5x tail tag at the N-terminal end of the proteins. The two AtSSIV versions were transformed into the E. coli strain BL21 (DE3) ΔglgCAP, which lacks the endogenous glycogen synthase activity. Selected clones containing the plasmids were cultivated and the expression of the AtSSIV versions were induced by IPTG. The levels of expression of the different versions were checked by immunoblot. One mL of each culture after the IPTG induction was collected, spun down in a microfuge and cells were resuspended in SDS-PAGE loading buffer and boiled for 10 min. The different samples were loaded into an SDS-PAGE gel; bands 45678were separated by electrophoresis and transferred to a ...
The old tricks could also be useful: lower the induction temperature and IPTG concentration, and shorten the induction time.... Does the degradation occur during induction or purification? Pete On Wed, 3 May 2000, Rick Thorne wrote: , Dear Eva , , All GST-fusions are different but I can offer you some guidelines which , might help you. , , First step might be to alter the strain of bacteria you are using for a , more specialized strain eg: BL21-DE3 from Stratagene (look at the web , site to learn more about these). Alternatives are available from other , companies (but I never used them). , , If this does not help, if at all practicable, make new construct/s with , smaller bits of the protein you want to use. This sometimes removes , parts of the molecule that encourage its wholesale destruction. , , regards, , , Rick , , Eva Chen wrote: , , , Is there anybody having experience on the purification of GST fusion , , proteins? , , Our lab doesnt have much experience and now there has been a ...
A model system has been developed to explore the relationship between cell cycle arrest and chemotherapeutic toxicity. An isopropyl-1-thio-β-d-galactopyranoside-inducible P16 construct was introduced stably into a melanoma cell line and used to promote G0-G1 arrest in the recipient cells. The state of arrest was reversible and did not compromise cell viability over a period of at least 7 days. Isopropyl-1-thio-β-d-galactopyranoside-treated, arrested cells were significantly more resistant to the chemotherapeutic agents methotrexate (∼50 times), vinblastine (,100 times), and cisplatin (∼10 times) compared to controls. This strategy of protection from chemotherapy exploits one of the basic genotypic differences between normal cells and tumor cells: the integrity of genetic pathways that regulate growth.. ...
Two polymorphic forms of bis[(E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]- pyrimidin-5-yl](3R,5S)-3,5-dihydroxyhept-6-enoic acid] calcium salt, processes for making them and t
This paper present ultraviolet-visible absorption spectra of imazamethabenz-methyl (IMBM) (mixture of the isomers methyl 6-[(RS)-4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl]-m-toluate, m-imazamethabenz, and methyl 2-[(RS)-4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl]-p-toluate, p-imazamethabenz) and the corresponding carboxylic acid, imazamethabenz-acid (IMBA). The spectral characteristics are determined as functions of the pH. The appreciable absorbance in the visible (or near-ultraviolet) region of the spectra indicates ...
The National Institute of Standards and Technology (NIST) uses its best efforts to deliver a high quality copy of the Database and to verify that the data contained therein have been selected on the basis of sound scientific judgment. However, NIST makes no warranties to that effect, and NIST shall not be liable for any damage that may result from errors or omissions in the Database ...
The National Institute of Standards and Technology (NIST) uses its best efforts to deliver a high quality copy of the Database and to verify that the data contained therein have been selected on the basis of sound scientific judgment. However, NIST makes no warranties to that effect, and NIST shall not be liable for any damage that may result from errors or omissions in the Database ...
小分子量熱休克蛋白質是植物體內種類最豐富的一群蛋白質,能與變性蛋白質結合,防止其沉澱而對細胞造成傷害。目前已知OsHSP16.9A與OsHSP18.0皆為水稻第一族群小分子量熱休克蛋白質的成員,分別位於第一和第三對染色體上,皆會在高溫41℃ ...
Gentaur molecular products has all kinds of products like :search , PhytoTechnology Laboratories \ IPTG \ I373-1G for more molecular products just contact us
polb cloned in pET not expressed - posted in Protein Expression and Purification: I have cloned a mutant human polb sequence in a pET28a vector. I have sequenced the clone, the sequence is in reading frame. When I try to expresses the protein in BL21 with IPTG, get no product. I have tried for several time, also in different concentration of IPTG, time period and temperature. Please help me.
Magenta-Gal is an X-Gal alternative that is used with IPTG to select successfully transformed cells. Magenta-Gal with IPTG is used in red-white screening.
TY - JOUR. T1 - Composite nanomaterials by self-assembly and controlled crystallization of poly(2-isopropyl-2-oxazoline)-grafted polysaccharides. AU - Morimoto, Nobuyuki. AU - Obeid, Rodolphe. AU - Yamane, Setsuko. AU - Winnik, Franoise M.. AU - Akiyoshi, Kazunari. PY - 2009/4/20. Y1 - 2009/4/20. N2 - We report here new composite nanomaterials by self-assembly and control of crystallization of thermoresponsive poly(2-isopropyl-2-oxazoline) (PIPOZ)-grafted pullulan.. AB - We report here new composite nanomaterials by self-assembly and control of crystallization of thermoresponsive poly(2-isopropyl-2-oxazoline) (PIPOZ)-grafted pullulan.. UR - http://www.scopus.com/inward/record.url?scp=64549101215&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=64549101215&partnerID=8YFLogxK. U2 - 10.1039/b817603e. DO - 10.1039/b817603e. M3 - Article. AN - SCOPUS:64549101215. VL - 5. SP - 1597. EP - 1600. JO - Soft Matter. JF - Soft Matter. SN - 1744-683X. IS - 8. ER - ...
It is shown that the 3-benzoyl-2-isopropyl-4-alkyloxazolidin-5-ones (12), derived from isobutyraldehyde and ?-amino acids are as efficient as, but more economical than, the corresponding tert-butyl compounds (9), as sources of enantiopure ?,?-dialkylated ?-amino acids (e.g., 21-23). In the absence of alkylating agents, the anions of (2R,4S)-12 and its enantiomer undergo a fragmentation-recombination process to generate (1S,2R,4S)-N-[1-(3-benzoyl- 2-isopropyl-4-methyl-5-oxo-oxazolidin-4-yl)-2-methylpropyl]benzamide ((1S, 2R,4S)-20), and its enantiomer. Acidic methanolysis of these condensation products provides access to ?,?-dialkylated ?,?-diaminopropionic acids [e.g., (2S,3S)-2,3-bisbenzoylamino-2,4-dimethylpentanoic acid methyl ester ((2S,3S)-24)). ...
Results. Metabolizable permissive glycan supports outer segment assembly: quantification of effect. Figure 1 illustrates examples of intact retinas that were removed from stage 33/34 Xenopus laevis tadpoles and placed into culture in Niu-Twitty medium for three days. Using this paradigm, all outer segment material is elaborated while in culture [29]. In retinas that were maintained with a normally apposed RPE, the outer segments are tightly stacked, properly folded and contain discs of equal diameter (Figure 1A). This morphology is identical to retinas maturing in vivo [29]. In RPE-deprived retinas that were otherwise similarly maintained, photoreceptor outer segment membranes are markedly disorganized, with little evidence of normal disc stacking (Figure 1B). The addition of 5x10-3 M mannose did not influence favorably the folding of outer segments in RPE-deprived retinas (Figure 1C), whereas the addition of 5x10-3 M lactose (Figure 1D) supported nicely the formation of nascent outer segments ...
Results show that in this condition both BBa_R0010 and BBa_R0011 produce different amounts of RFP as a function of the IPTG concentration. The amplitude of the two curves show that the promoters are very strong when induced with IPTG ,= 10 uM. Although the experiments were carried out in the same conditions, the variability between experiments was high, especially for BBa_R0010 (mean coefficient of variaton of about 37% for BBa_R0010 and 15% for BBa_R0011), while the RPU variability between three wells in the same experiment is much lower (mean coefficient of variaton of bout 3.5% for both promoters). The above figure shows that BBa_R0011 is stronger than the BBa_R0010 wild type promoter in low copy plasmid. This result is unexpected because the same promoters in high copy vectors behaved differently (BBa_R0010 was stronger than the BBa_R0011, see above). In the uninduced state, BBa_R0011 has about the same strength as the BBa_J23101 reference standard promoter. This static characteristic shows ...
2-isopropyl-5,5-dimethyl-5,6-dihydro-2H-1,3-oxazine - chemical structural formula, chemical names, chemical properties, synthesis references
4-chloro-2-isopropyl-6-methylpyrimidine 1-oxide - chemical structural formula, chemical names, chemical properties, synthesis references
(2E,4E,6Z)-7-(5-Isopropyl-3-pentafluoroethyl-2-propoxy-phenyl)-3-methyl-octa-2,4,6-trienoic acid | C23H27F5O3 | CID 44353456 - structure, chemical names, physical and chemical properties, classification, patents, literature, biological activities, safety/hazards/toxicity information, supplier lists, and more.
6-{[2-(2-Isopropyl-4-methylphenoxy)ethyl]thio}-9H-purine | C17H20N4OS | CID 2257073 - structure, chemical names, physical and chemical properties, classification, patents, literature, biological activities, safety/hazards/toxicity information, supplier lists, and more.
40853-56-3 - AOWLXZIJUIFJPM-KPKJPENVSA-N - Acetic acid, 2-isopropyl-5-methyl-2-hexen-1-yl ester - Similar structures search, synonyms, formulas, resource links, and other chemical information.
10-isopropyl-2,2,6-trimethyl-2,3,4,5-tetrahydronaphtha(1,8-bc)oxocine-5,11-diol: from the roots and rhizomes of Nardostachys chinensis; structure in first source
The use of λ repressor fusions to study protein-protein interactions in E. coli was first described by Hu and others (1). Since then, the repressor system has been employed by several laboratories to...
Global transcription responses of Escherichia coli to various stimuli or genetic defects were studied by measuring mRNA levels in about 400 segments of the genome. Measuring mRNA levels was done by analyzing hybridization to DNA dot blots made with overlapping lambda clones spanning the genome of E. coli K-12. Conditions examined included isopropyl-beta-D-thiogalactopyranoside (IPTG) induction, heat shock, osmotic shock, starvation for various nutrients, entrance of cells into the stationary phase of growth, anaerobic growth in a tube, growth in the gnotobiotic mouse gut, and effects of pleiotropic mutations rpoH, himA, topA, and crp. Most mapped genes known to be regulated by a particular situation were successfully detected. In addition, many chromosomal regions containing no previously known regulated genes were discovered that responded to various stimuli. This new method for studying globally regulated genetic systems in E. coli combines detection, cloning, and physical mapping of a battery ...
Inducible expression systems are frequently used for the production of heterologous proteins. Achieving maximum product concentrations requires induction profiling, namely the optimization of induction time and inducer concentration. However, the respective experiments can be very laborious and time-consuming. In this work, a new approach for induction profiling is presented where induction in a microtiter plate based cultivation system (BioLector) is achieved by light using photocaged isopropyl β-d-1-thiogalactopyranoside (cIPTG). A flavin mononucleotide-based fluorescent reporter protein (FbFP) was expressed using a T7-RNA-polymerase dependent E. coli expression system which required IPTG as inducer. High power UV-A irradiation was directed into a microtiter plate by light-emitting diodes placed above each well of a 48-well plate. Upon UV irradiation, IPTG is released (uncaged) and induces product formation. IPTG uncaging, formation of the fluorescent reporter protein and biomass growth were
For years I have been teaching my students that a gene is a segment of DNA that codes for a single RNA molecule with a complementary sequence, regardless of whether that RNA molecule is translated or not. This definition takes into account the genes for the various rRNAs and tRNAs, which are not translated, and also other forms of non-translated RNA that have recently been discovered. By this definition, genes that code for mRNAs that are actually translated are distinguished as structural genes, using terminology that was first developed to describe the Jacob-Monod model of the lactose operon. Using this same terminology, the gene that codes for the lactose repressor protein is a regulatory gene, insofar as the repressor does not function in an extrinsic biochemical pathway, but rather participates in the regulation of other structural genes. However, the distinction between structural and regulatory genes outlined above is insufficient to describe the various kinds of genetically ...
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The RE Family based on Renesas SOTB™ process technology realizes ultra-low current consumption in active and standby modes and fast operation at low voltage.
Mouse polyclonal antibody raised against a partial recombinant CHERP. CHERP (AAH21294.1, 795 a.a. ~ 883 a.a) partial recombinant protein with GST tag. (H00010523-A01) - Products - Abnova
Nitrogen › N-[(1R,2R)-2-(Dimethylamino)-1,2-diphenylethyl]-N-[[(1R,4aS,10aR)-1,2,3,4,4a,9,10,10a-octahydro-1,4a-dimethyl-7-isopropyl-1-phenanthrenyl]methyl]thiourea, 98%, (99% ee) ...
5-(4-ethylpiperazin-1-ylmethyl)pyridin-2-yl)-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-3H-benzimidazol-5-yl)pyrimidin-2-yl)amine ...
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
4-ISOPROPYL-2-PHENYL-2-OXAZOLINE-5-ONE (CAS 5839-93-0) Market Research Report 2018 aims at providing comprehensive data on 4-isopropyl-2-phenyl-2-oxazoline-5-one
Thymol; p-Cymen-3-ol; Thyme camphor; 2-Isopropyl-5-methylphenol; 3-Hydroxy-p-cymene; 3-Methyl-6-isopropylphenol; 5-Methyl-2-isopropylphenol; 6-Isopropyl-m-cresol; 6-Isopropyl-3-methylphenol; m-Cresol, 6-isopropyl-; p-Cymene, 3-hydroxy-; Isopropyl cresol; Phenol, 2-isopropyl-5-methyl-; Thymic acid; 3-p-Cymenol; 5-Methyl-2-isopropyl-1-phenol; 5-Methyl-2-(1-methylethyl)phenol; Isopropyl-m-cresol; m- ...
This page contains information on the chemical Barbituric acid, 5-(2-bromoallyl)-5-isopropyl-1-methyl-, sodium salt including: 11 synonyms/identifiers.
64038-35-3 - ZOKRAMSWASSLAJ-IPZCTEOASA-M - Barbituric acid, 5-isopropyl-5-(2-pentenyl)-, sodium salt - Similar structures search, synonyms, formulas, resource links, and other chemical information.
View Notes - 351 E2p mc from CHEM 351 at BYU. 351 E2 Practice 1. How many stereoisomers are there of 1-isopropyl-4-methylcyclohexane? A. only 1 structure possible - no stereoisomers B. two C. three
This research project attempted to grow and purify soluble Pseudomonas putida HMGR (PpHMGR) enzyme for the purpose of experimentation on its hypothesized morpheein protein behavior. Inclusion body formation has been the primary limiting factor in the growth and purification of soluble PpHMGR. Variation on incubation temperature, IPTG concentration, and growth time were explored in the growth phase, and imidazole concentration, presence of DTT, and presence of TCEP were explored in the purification phase. PpHMGR was grown in an E. coli host, cell lysis was performed by sonication, and subsequent purification was performed by nickel column elution FPLC. The purified product was characterized by FPLC, gel electrophoresis, and enzyme kinetics to determine the reduction of inclusion body formation and presence of soluble PpHMGR.
Supplementary MaterialsAdditional file 1: Figure S1. expression of the TetR family protein when induced with different inducer concentrations. Lane1: marker, lane2: cells with uninduced TetR family protein, lane3-6: cells with induced TetR family protein (0.2?mM, 0.5?mM, 1?mM and 2?mM IPTG concentration). 13568_2019_801_MOESM1_ESM.pdf (426K) GUID:?E4D189AF-6495-44A5-9CF1-3575D2773CCA Data Availability StatementAll data analyzed throughout this Mouse monoclonal to EhpB1 study is shown in the article. Abstract Biodesulfurization helps in removal of sulfur from organosulfur present in petroleum fractions. All microorganisms isolated to date harbor a desulfurization operon consisting of three genes and -which encode for monooxygenases (DszA & C) and desulfinase (DszB). Most of the studies have been carried out using dibenzothiophene as the model organosulfur compound, which is converted into 2 hydroxybiphenyl by a 4S pathway which maintains the calorific value of fuel. There are few studies reported ...
A putative operon encoding a probable zinc-responsive regulatory element (zur) and components of an ABC-type transporter (mreA mreB) have been characterized in Staphylococcus aureus. The zur gene was inactivated but apparently this did not alter Zn2+ uptake. Expression of mreAB zur is at a low level under a range of ion conditions. To allow inducible expression of the operon, a construct was made placing it under the control of the IPTG-inducible Pspac promoter. Using this approach, it was shown that zur is able to repress expression of the entire operon in a Zn2+-dependent manner, and that mreA and mreB are likely to be involved in high-affinity ion uptake. zur has no apparent role in pathogenicity in a lesion model of S. aureus infection.
The Argonaute protein of Thermus thermophilus (TtAgo) has recently been studied in detail. For its in vitro characterization, TtAgo was purified after heterologous expression in Escherichia coli (E. coli). As TtAgo expression is toxic, a tightly controlled system was used for protein expression. The expression strain E. coli KRX carries a chromosomal T7 RNA polymerase gene under control of a rhamnose promoter. The ago gene is expressed via an IPTG-inducible T7 promoter. This allows for tightly (double) controlled expression of (toxic) TtAgo. Here, we describe the steps required for controlled expression and purification of this toxic protein.
More specifically: The bacterial cells of strain HTll5(DE3) contain the T7 RNA polymerase gene (contained within a stable insertion of a modified lambda prophage λ DE3) under the control of lac operon regulatory elements. This allows expression of T7 polymerase to be controlled by isopropyl-β-D-thiogalactopyranoside (IPTG), a lactose analogue that induces expression of genes under the control of the lac operon o gene. When IPTG is added, the cells will begin to synthesize lots of T7 RNA polymerase. This T7 RNA polymerase can then bind to the T7 promoter sites on the plasmid and begin to synthesize RNA from both T7 RNA polymerase sites. Because the two single strands of RNA are complementary to each other they will form double stranded RNA within the bacterial cell. The IPTG induction allows us to turn on and express the plasmid gene of interest only when we want to and it allow us to make much higher levels of RNA for RNA interference than would be made without this induction. Another useful ...
More specifically: The bacterial cells of strain HTll5(DE3) contain the T7 RNA polymerase gene (contained within a stable insertion of a modified lambda prophage λ DE3) under the control of lac operon regulatory elements. This allows expression of T7 polymerase to be controlled by isopropyl-β-D-thiogalactopyranoside (IPTG), a lactose analogue that induces expression of genes under the control of the lac operon o gene. When IPTG is added, the cells will begin to synthesize lots of T7 RNA polymerase. This T7 RNA polymerase can then bind to the T7 promoter sites on the plasmid and begin to synthesize RNA from both T7 RNA polymerase sites. Because the two single strands of RNA are complementary to each other they will form double stranded RNA within the bacterial cell. The IPTG induction allows us to turn on and express the plasmid gene of interest only when we want to and it allow us to make much higher levels of RNA for RNA interference than would be made without this induction. Another useful ...
When using this server please cite the following paper:. Zsila F, Bikadi Z, Malik D, Hari P, Pechan I, Berces A, Hazai E.. Evaluation of drug-human serum albumin binding interactions with support vector machine aided online automated docking.. Bioinformatics. 2011 May 18. ...
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Chemical Entities of Biological Interest (ChEBI) is a freely available dictionary of molecular entities focused on small chemical compounds.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice ...
The RE family based on the Silicon on Thin Buried Oxide (SOTB™ ) process technology realizes both ultra-low current consumption in both active and standby mode and high speed CPU operation (64MHz) at low voltage (1.62V) , which is impossible to achieve wi...
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The present invention relates to 4-{(1R,3R)-1-(3,5-difluorophenyl)-3-[4-(3-ethyl-5-isopropyl-4H-1,2,4-triazol-4-yl)piperidin-1-yl]butyl}-1-(methylsulfonyl)piperidine (I): |p||chemistry id=CHEM-US-000
Torsemide is a diuretic of the pyridine-sulfonylurea class. Its chemical name is 1-isopropyl-3-[(4-m-toluidino-3-pyridyl) sulfonyl] urea. Torsemide acts from within the lumen of the thick ascending portion of the loop of Henle, where it inhibits the Na/K+/2CI- carrier system. ...
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