Biotin switch assay
WebThe biotin switch assay, developed by Jaffrey et al., is a common method of detecting SNOs, and the steps of the assay are listed below: All free cysteines are blocked. All remaining cysteines (presumably only those that are denitrosylated) are denitrosylated. WebThe biotin-switch assay was then performed as described above. Quantification and statistical analysis. Fiji (ImageJ) and Image Studio were used for quantification of immunohistochemistry and immunoblot data, respectively. A Power Analysis of our prior data was used to determine the number of replicates needed for statistical purposes.
Biotin switch assay
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WebSep 22, 2009 · Endogenous S-nitrosylation of stargazin. (A) HEK cells expressing WT or C302S mutant of stargazin were treated with Cys-NO (50 μM) for 5–10 min, and analyzed by the biotin-switch assay.(B) The quantity of SNO-stargazin from A was measured by scanning densitometry, and expressed as percentage WT SNO-stargazin.Data are … WebSep 28, 2024 · H: REDD1 oxidation was evaluated by biotin switch assay. PKM2 was evaluated in biotin pull-downs as a positive control. HA-REDD1 and PKM2 expression in whole-cell lysate (WCL) was determined by Western blotting. I: WT and REDD1-KO MIO-M1 cells were exposed to medium containing 30 mmol/L glucose (HG) for up to 6 h, and …
WebMar 14, 2024 · The biotin-switch assay showed that BR treatment clearly induced the oxidation of BZR1, and application of H 2 O 2 enhanced the oxidative effect ... WebAug 19, 2013 · In 1998, Jaffrey et al. described the biotin switch assay, which used HPDP-biotin (Part No. 21341) to detect S-nitrosylated proteins (Ref.2). In this assay, free sulfhydryls are chemically blocked, whereas nitrosylated cysteines are selectively reduced and biotinylated for streptavidin-based enrichment and/or detection. However, the biotin ...
WebMar 13, 2024 · Biotin-switch assay was performed on WT, C207H and C279H TH expressed in HEK293T cells, C279H TH was found to have a reduction on TH S-nitrosylation. ( B ) WT and C279H TH were subjected to real ... WebJun 12, 2001 · Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. Here, we describe a novel method for detecting proteins that contain nitrosothiols. In this three-step procedure, nitrosylated cysteines are converted to bio …
WebNov 10, 2009 · Fig. 1 H 2 S covalently modifies proteins through S-sulfhydration of cysteine residues. (A) Liver lysates treated with 100 μM NaHS for 30 min at 37°C and subjected to the modified biotin switch assay with antibody against biotin (Anti-biotin Ab) to detect S-sulfhydration show numerous sulfhydrated proteins.(B) LC-MS/MS of a subset of the …
WebJan 1, 2009 · The focus of the current review is the biotin switch technique (BST), which has become a mainstay assay for detecting S-nitrosylated proteins in complex biological … razer tron mouseWebProduct overview. Biotin Switch Assay Kit (S-Nitrosylation) (ab236207) uses a modification of the Jaffrey et al. 'Biotin-switch' method to measure S-Nitrosylated (S … simpson pharmacy 1015 southern blvdWebDec 1, 2024 · A warning for people who take the dietary supplement biotin (vitamin B7): the FDA says taking too much of the vitamin may interfere with the results of some lab tests. … razer tron keyboard soundsWebThe standard biotin switch assay was used to reveal the presence of NO-induced modifications in GSNO-treated HPAEC lysates (Fig. 2A). Silver stain gel analysis shows a clear increase in the amount of protein … simpson ph612i screwsWebBiotin quantitation kits are used for determining biotinylation levels and efficiency by either colorimetric or fluorescence detection methods. These kits contain the simple and … razer true wireless appWebMar 1, 2004 · Biotin switch assay, Western blot, and protein purification The biotin switch assay was performed essentially as previously described [22] , [42] . Extracts were adjusted to 0.5 mg/ml of protein and equal amounts were blocked with 4 volumes of blocking buffer (225 mM Hepes, pH 7.7, 0.9 mM EDTA, 0.09 mM neocuproine, 2.5% SDS, and 20 mM … simpson pharmacy bronxWebJun 12, 2001 · Purification of biotinylated proteins. 1. To the samples from "Biotinylation of nitrosothiols," add two volumes of -20°C acetone and incubate for 20 min at -20°C to … simpson pharmacy galgorm