Sigma
Anti-Glucose-6-Phosphate Dehydrogenase (G-6-PDH) antibody produced in rabbit IgG fraction of antiserum, lyophilized powder
A9521
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The aim of this experiment was to study the expression dynamics of budding yeast G1 cyclin Cln2 after release from a G1 block caused by pheromone alpha-factor. G6PDH antibody is used here as a loading control and it works fabulously.
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-Rabbit-HRP, 1:3000, 1hour incubation at Room Temperature
None
Biorad Clarity HRP substrate at 1:10 dilution
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 then 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 at 1:10 dilution. The antibody detected endogenous levels of G6PDH very strongly.
Great antibody for G6PDH
I would use it for detecting G6PDH in a variety of cell types