Strep-Tactin®与Strep-Tactin®XT纯化流程的差异主要在于: 将靶蛋白从Strep-Tactin®XT洗脱时,需使用含有50 mM Biotin (生物素)的Buffer BXT (由于亲和力大幅提升,Desthiobiotin不能有效的洗脱靶蛋白。) 再生Strep-Tactin®XT Superflow®填料时,需使用3 M...
After incubation of the lysate with the beads, the column was washed with wash buffer containing 50 mm Tris-HCl, pH 8.0, 200 mm NaCl, and protein was eluted with wash buffer containing 3 mm d-desthiobiotin (iba-2-1000-005). The elution pool was concentrated using a Vivaspin 10,000 ...
Triton X100, Nonidet P-40 (NP40),imidazole, ATP, glycerol, desthiobiotin. 7. Buffer A: phosphate-buffered saline (PBS) pH 8.0 containing 20% glycerol, 1 mM (2-carboxyethyl)phosphine·HCl (TCEP), and EDTA-freeprotease inhibitorcocktail (Bimake). ...
(GE healthcare). Proteins were eluted from the column with 2.5 mM desthiobiotin and subsequently loaded on and eluted from a Superdex 200 (GE healthcare), equilibrated and run with 50 mM TrispH 8, 150 mM NaCl and 5 mM MgCl2. Selenium labelled proteins were transformed in T7 ...
将靶蛋白从Strep-Tactin®XT洗脱时,需使用含有50 mM Biotin (生物素)的Buffer BXT (由于亲和力大幅提升,Desthiobiotin不能有效的洗脱靶蛋白。) 再生Strep-Tactin®XT填料时,需使用3 M MgCl2 (因使用生物素进行洗脱,HABA无法与生物素进行竞争,需以MgCl2进行再生...