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 … WebFeb 21, 2012 · The biotin switch assay followed by Western blot or immunoprecipitation using an S-nitrosocysteine antibody was also employed to validate S-nitrosylated proteins in endothelial cell lysates. Results Seventy-eight potential S-nitrosylated proteins and their target cysteine residues for S-nitrosylation were identified; 9 of them were Golgi ...
H2S Signals Through Protein S-Sulfhydration Science Signaling
WebJan 11, 2008 · The biotin switch assay uses N-ethylmaleimide (NEM) to block free sulfhydryls followed by cleavage of thioesters with hydroxylamine and labelling of liberated sulfhydryls with biotin (Figure (Figure1). 1). In the negative controls sulfhydryl reactive biotin is still present but Tris, which does not cleave thioester bonds, replaces … WebSep 9, 2009 · Cell extracts were analyzed by the biotin-switch assay as described before with modifications . Briefly, cells were extracted in HEN buffer (250 mM Hepes/1 mM EDTA/0.1 mM Neocuproine, pH 7.7) containing Triton X-100 (1%). Extracts were treated with methyl methanethiosulfonate (Sigma–Aldrich) and SDS (1%) at 50 °C for 20 min. … the westin annapolis spa
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WebJun 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 … WebCayman Chemical biotin switch Biotin Switch, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS … WebDec 2, 2016 · Using arsenite as reducing agent in the modified biotin switch assay, we were able to quantify 1161 proteins, in which more than 100 sulfenic acid sites were identified. Bioinformatics analysis of the quantified sulfenic acid sites highlighted the relevant, previously missed biological process of monocyte transendothelial migration, … the westin annapolis parking