Author: Rathore, Shailendra S.; Liu, Yinghui; Yu, Haijia; Wan, Chun; Lee, MyeongSeon; Yin, Qian; Stowell, Michael H.B.; Shen, Jingshi
Title: Intracellular Vesicle Fusion Requires a Membrane-Destabilizing Peptide Located at the Juxtamembrane Region of the v-SNARE Document date: 2019_12_24
ID: pudp1eoo_33
Snippet: Microbial Strains-All the recombinant proteins in this study were expressed in E. Coli BL21 [B F − ompT hsdS(r B Vc peptide (residues 56-84 of VAMP2) were expressed and purified in E. coli using the pET28a vector Yu et al.,2018) . SNARE mutants were prepared similarly as the corresponding WT proteins. Full-length SNAREs were stored in a buffer containing 25 mM HEPES (pH 7.4), 400 mM KCl, 1% n-octyl-β-D-glucoside (OG), 10% glycerol, and 0.5 mM .....
Document: Microbial Strains-All the recombinant proteins in this study were expressed in E. Coli BL21 [B F − ompT hsdS(r B Vc peptide (residues 56-84 of VAMP2) were expressed and purified in E. coli using the pET28a vector Yu et al.,2018) . SNARE mutants were prepared similarly as the corresponding WT proteins. Full-length SNAREs were stored in a buffer containing 25 mM HEPES (pH 7.4), 400 mM KCl, 1% n-octyl-β-D-glucoside (OG), 10% glycerol, and 0.5 mM Tris(2-carboxyethyl)phosphine (TCEP). Soluble proteins were stored in a protein binding buffer (25 mM HEPES [pH 7.4], 150 mM KCl, 10% glycerol, and 0.5 mM TCEP).
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