Anti-Raf1 (phospho Ser621) Antibody (A10884)

Western blot analysis of extracts of HeLa and 293T cells, using Anti-Raf1 (phospho Ser621) Antibody (A10884) at 1:1,000 dilution. HeLa cells were treated by EGF (100ng/ml) for 30 minutes after serum-starvation overnight. 293T cells were treated by PMA/TPA (200nM) for 30 minutes or treated by EGF (25ug/mL) for 30 minutes after serum-starvation overnight. The secondary antibody was Goat Anti-Rabbit IgG H&L Antibody (HRP) at 1:10,000 dilution. Lysates/proteins were present at 25µg per lane. The blocking buffer used was 3% BSA.

Immunoprecipitation analysis of 200µg extracts of HeLa cells using 3µg of Anti-Raf1 (phospho Ser621) Antibody (A10884). This Western blot was performed on the immunoprecipitate using Anti-Raf1 (phospho Ser621) Antibody (A10884) at a dilution of 1:1000. HeLa cells were treated by EGF (100 ng/ml) at 37°C for 30 minutes after serum-starvation overnight.