Anti-Hsp27 (phospho Ser82) Antibody (A16949)

Western blot analysis of extracts of HeLa cells, using Anti-Hsp27 (phospho Ser82) Antibody (A16949) at 1:1,000 dilution. HeLa cells were treated by EGF (100ng/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. Detection was with a ECL Basic Kit. Exposure time: 1min.

Western blot analysis of extracts of HeLa cells, using Anti-Hsp27 (phospho Ser82) Antibody (A16949) at 1:1,000 dilution or HSP27/HSPB1 antibody (A16332). HeLa cells were treated by UV at room temperature for 15-30 minutes. HeLa cells were treated by Anisomycin (25 ug/mL) at 37°C 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. Detection was with a ECL Basic Kit. Exposure time: 10s.

Immunoprecipitation analysis of 200µg extracts of HeLa cells using 3µg of Anti-Hsp27 (phospho Ser82) Antibody (A16949). This Western blot was performed on the immunoprecipitate using Anti-Hsp27 (phospho Ser82) Antibody (A16949) 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.