Anti-AKT1 (phospho Ser473) Antibody (A90652)

Western blot analysis of 293F, using Anti-AKT1 (phospho Ser473) Antibody (A90652) at 1:1,000 dilution. 293F cells were treated by IGF-1 (50 ng/ml) at 37°C for 5 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% non-fat dry milk in TBST. Detection was with a ECL Basic Kit. Exposure time: 1s.

Western blot analysis of NIH/3T3, using Anti-AKT1 (phospho Ser473) Antibody (A90652) at 1:1,000 dilution. NIH/3T3 cells were treated by PDGF (100 ng/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% non-fat dry milk in TBST. Detection was with a ECL Basic Kit. Exposure time: 1s.

Immunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue using Anti-AKT1 (phospho Ser473) Antibody (A90652) at a dilution of 1:50 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunoprecipitation analysis of 200µg extracts of Jurkat cells using 3µg of Anti-AKT1 (phospho Ser473) Antibody (A90652). This Western blot was performed on the immunoprecipitate using Anti-AKT1 (phospho Ser473) Antibody (A90652) at a dilution of 1:1000. Jurkat cells were treated by Calycµlin A (100 nM) at 37°C for 30 minutes.