Akt Isoform Specific Antibody Sampler Kit provides an economical means of distinguishing between the three Akt isoforms, Akt1, Akt2 and Akt3, as well as a pan-Akt antibody. The kit provides enough primary and secondary antibodies to perform two Western blot experiments.
Specificity / Sensitivity
Each isoform-specific antibody recognizes endogenous levels of the specified Akt isoform independent of its phosphorylation state. Neither antibody cross-reacts with the two other isoforms. Akt Antibody recognizes endogenous levels of all three Akt isoforms.
Source / Purification
Each isoform-specific antibody is produced by immunizing mice or rabbits with a synthetic peptide that is specific for the Akt isoform. Akt1 Monoclonal antibody is produced by immunizing animals with a synthetic peptide surrounding Leu110 of human Akt1. Akt2 monoclonal antibody #3063 is produced by immunizing rabbits with a synthetic peptide corresponding to residues of human Akt2. Akt3 monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg116 of human Akt3 protein. Akt(pan) monoclonal antibody is produced by immunizing animals with a synthetic peptide at the carboxy-terminal sequence of mouse Akt.
Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3K/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip1 (15) and p21 Waf1/Cip1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).