Biotin-switch assay
WebJan 1, 2010 · Both biotin switch assay and sinapinic acid technique are used for detecting S-nitroso-BSA (Fig. 6, Lanes 3 and 6). Download : Download full-size image; Fig. 5. … 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 ...
Biotin-switch assay
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WebClassic biotin-switch assay and its derivatives are the most common methods used for detecting SNO. In this approach, the labile SNO group is selectively replaced with a … WebFor quantitative determination of biotin in food, cosmetic, and biotinylated proteins. Key Features. Fast and sensitive. Linear detection range: 8 to 200 µM biotin with 30 µL sample (96-well) or 10 to 200 µM biotin with 10 µL sample (384-well). Convenient. The procedure involves adding a single working reagent and reading after 20 minutes.
WebS-nitrosylation (or S-nitrosation, SNO) is an oxidative posttranslational modification to the thiol group of a cysteine amino acid residue. There are several methods to detect SNO modifications, mostly based on the classic biotin-switch assay, where the labile SNO sites are replaced with a stable biotin moiety to facilitate enrichment of the modified proteins. WebJan 1, 2024 · The biotin switch assay has been widely applied to the quantitative analysis of S-nitrosylation in different disease models and is now also emerging as a valuable research tool for other oxidative cysteine modifications, highlighting its relevance as a versatile, robust strategy for carrying out in-depth studies in redox proteomics.
WebCayman’s S- Nitrosylated Protein Detection Kit (Biotin Switch) employs a modification of the Jaffrey et al. 'Biotin- switch' method to allow for the direct visualization of S- NO … WebJan 1, 2010 · Both biotin switch assay and sinapinic acid technique are used for detecting S-nitroso-BSA (Fig. 6, Lanes 3 and 6). Download : Download full-size image; Fig. 5. Western blot of protein S-nitrosothiols in control and LPS-treated with and without l-NMMA in RAW 264.7 cells detected by both sinapinic acid assay and biotin switch assay.
WebFeb 1, 2024 · Biotin-switch assay of protein S-sulfhydration S-sulfhydration was detected by the biotin-switch method using the purified BraFLC protein treated with H 2 S. The method was performed as previously ...
WebThe assay is a variation of the modified biotin switch assay. The reaction mixture is treated with the thiol blocking reagent (BR): methylsulfonyl benzothiazole (MSBT), followed by treatment with a methylcyanoacetate (MCA) derivative that comprises a nucleophilic component and a biotin moiety as a reporter. css flex stretchWebArticle Title: Copper dependence of the biotin switch assay doi: 10.1016/j.freeradbiomed.2007.12.032 Figure Lengend Snippet: Equal amounts of HSA-SNO and HSA were subjected to the biotin switch assay. earl charles grey black butlerWebJan 2, 2015 · Random (−) or CBS (+) siRNA-treated cells were incubated with 1 mM l-NAME for 16 h, and then assayed using the biotin switch method. Right panel: biotin-switch assay and Western analysis were quantified; the relative ratio of SNO-PTEN to total PTEN was calculated for each sample. Values are means ± S.E., n = 3. ∗∗ p < 0.01 by ANOVA. css flex table rowsWebFeb 4, 2013 · For the biotin-switch assay, cortical hemispheres (n = 3) were harvested from the control or ischemic side of the brain immediately after reperfusion. SHP-2 PTP Activity Assay. In vitro phosphatase activity of recombinant Shp-2 with pNPP (Sigma) as a substrate was measured in a 30-μL reaction mixture containing 20 mM pNPP, 50 mM … css flex syntaxWebClassic biotin-switch assay and its derivatives are the most common methods used for detecting SNO. In this approach, the labile SNO group is selectively replaced with a single stable tag. To date, a variety of thiol-reactive tags have been introduced. However, these methods have not produced a consistent data set, which suggests an incomplete ... earl charlesWebThe biotin switch assay was performed as previously described (Hemsley, Taylor, & Grierson, 2008) Hydroxylamine was replaced by Tris-HCl buffer in the remaining aliquot … css flex-startWebJan 10, 2024 · Here, we report an adapted biotin switch assay method applying to the protein persulfidation level detection. As shown in Fig. 1 a, b, the purified recombinant … css flex tabs