Abcam
Anti-beta Actin antibody
ab8227
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Beta-actin is often used as a loading control for Western Blotting. It is a cytoskeleton protein, present in all eukaryotic cells. We used this particular antibody to probe internal Beta actin in Hela cells.
Western Blot
HeLa cell lysates
1:500 dilution prepared in blocking solution, incubated overnight at 4 degrees
5% skimmed non-fat milk in PBST20
1:1000 anti-rabbit HRP antibody, incubated for an hour at RT
N/A
ECL based detection was used
We observed reduced phosphorylation upon treatment in Hela cells. There were few non-specific bands but prominent band was of phospho Akt
Good antibody
None
Overall, it is a good antibody to detect phosphorylation of Akt.