SDS-PAGE and Western blot analysis
The protein sample prepared from empty vector transfected cells,
uninduced cells, induced cells, purified pB602L or polypeptides was
mixed with loading buffer and boiled for 10 min. The proteins in each
sample were separated by 10% gel and stained with Coomassie Brilliant
blue R-250 for SDS-PAGE. For Western blot analysis, the gel was blotted
to PVDF transfer membrane with an electrophoresis apparatus. The
membrane was blocked by 5% Difco Skim Milk for 2 h at room temperature
and then incubated with each of the mAbs or a mouse monoclonal anti-GST
antibody (Sigma-Aldrich, USA) as primary antibody at room temperature
for 1 h. After four washes with Tris-Buffered Saline (TBS) add 0.5%
Tween-20 (TBST), the membrane was incubated with IRDye 800CW goat
anti-Mouse IgG (H + L) as secondary antibody (LICOR, USA) at room
temperature for 1 h, followed by four washes with TBST. Western blot
analysis was completed with an Odyssey CLX (LICOR, USA).