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AKT, also known as protein kinase B (PKB) or Related to A and C kinases (RAC), is the cellular homolog of the viral oncogene v-AKT. It was originally cloned based on its homology to protein kinases A and C. AKT is activated in response to insulin, and various growth and survival factors. It is clear that AKT enzyme activity is both necessary and essential for cell survival, and AKT has been shown to be able to induce progression of the cell cycle as well as suppress apoptosis. The key role of AKT in the regulation of cell survival makes it a promising target for anti-oncogenic therapeutics.
Figure 1: Measurement of phosphorylated and total AKT.
NIH/3T3 cells were cultured in 96-well plates and serum-starved for 16 hours. Cells were then treated with 50 ng/ml PDGF for 5 minutes and fixed. Total and phospho-AKT were each assayed in triplicate using the phospho and total AKT antibodies from the FACE AKT Kit.
Antibody Specificities
The phospho-AKT antibody is specific for phosphorylated AKT and was raised against a phospho-Ser473 peptide that corresponds to the sequence surrounding Ser473 of mouse AKT. It recognizes AKT1 only when phosphorylated at Ser473, and AKT2 and AKT3 only when phosphorylated at equivalent sites. The total-AKT antibody recognizes AKT1, AKT2 and AKT3 regardless of the phosphorylation state.
