Cell Signaling Technology
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb
4060
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The aim of our work is to check the levels of AKT phosphorylation at serine 473 (pAKT ser473) in TSC2-knockdown cells to understand the regulation of mTORC2 signaling by TSC2. For that, we analyzed the levels of phosphorylation of AKT at serine473 in scramble and siRNA-mediated TSC2-knockdown in HeLa whole cell lysates.
Western Blot
HeLa whole cell lysate
1:3000, overnight at 4°c
5% skimmed milk, 1 hr at room temperature
1:10000, anti-rabbit HRP conjugated antibody, 1 hr at room temperature
N/A
ECL reagent, Chemi doc
We observed a single band corresponding to pAKT at 60KD in western blot. As shown in the literature, we observed decreased levels of pAKT in TSC2-knockdown cell lysates compared to scramble.
29143563
Excellent antibody and easy to optimize for performing western blots
Gives a single specific band on western blot and easy to optimize.