Santa Cruz Biotechnology
GST Antibody (B-14): sc-138
sc-138
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The aim of this experiment was to study the function of a variety of yeast G1 cyclins by expressing them in budding yeast Saccharomyces cerevisiae as C-terminal GST-tagged proteins and using a common substrate. Here the exression of GST tagged cyclins are detected successfully
Western Blot
Cell Lysates from budding yeast
1:1000 Dilution in 5% Milk (TTBS), overnight incubation in cold room
5% non-fat milk in TBS-T
Anti-Mouse-HRP, 1:3000, 1hour incubation at Room Temperature
None
Bio-Rad Clarity HRP substrate
Protein extract from cells was prepared in TCA lysis buffer supplied with PMSF, protease inhibitor mixture, and sodium orthovanadate. Lysates were quantified using a BCA assay kit (Thermos). 20 micrograms of proteins were subjected to SDS-PAGE and transferred onto PVDF membranes and blocked with 5% non-fat milk in TBS-T (20 mMTris, 500 mMNaCl, and 0.1% Tween 20) at room temperature for 1 h with rocking. The membranes were probed with primary Antibody (1/1000 dilution) overnight at 4°C. After washing with TBS-T, the membranes were incubated with HRP-conjugated secondary antibody in 5% non-fat milk/TBS-T at room temperature for 1 h. The protein–Antibody complex was detected by Bio-Rad Clarity ECL substrate. The antibody detected GST-tagged cyclins robustly.
Works great for GST
I would use this antibody for detection of GST in any cells.