Rabbit Polyclonal to Akt1 / PKB
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Serine/threonine protein kinase Akt/PKB
(Protein Kinase B) plays important roles in cell growth, metabolism, proliferation and survival. Upon extracellular stimulus that leads to activation of phosphatidylinositol 3-kinase (PI3K), Akt is recruited from the cytosol to the plasma membrane through binding 3´phosphoinositides and is phosphorylated. Active Akt dissociates from the plasma membrane to gain access to various substrates in different subcellular locations by translocating to cytosol and nucleus.
The polyclonal antibody reacts with Akt1 (recognized epitope: between aa 85-100 of Akt1/PKB). Akt/PKB protein kinases family has a three mammalian members identified to date (Akt1, Akt2, Akt3). Human Akt1 has a predicted molecular weight of 55.7 kDa.
Synthetic peptide (coupled with KLH) derived from aminoacid sequence 85-100 of human Akt1. This immunogen sequence has 100% homology with mouse and rat Akt1/PKB.
- Western Blotting
Recommended dilution: 0.5-1 μg/ml
Positive material: RAJI human Burkitt lymphoma cell line
Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing/reducing Laemmli SDS-PAGE sample buffer. Boil for 5 min.
Application note: Both reducing and non-reducing conditions. Reducing conditions are recommended.
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.
*Ananthanarayanan B, Fosbrink M, Rahdar M, Zhang J: Live-cell molecular analysis of AKT activation reveals roles for activation loop phosphorylation. J Biol Chem. 2007 Oct 10. [Abstract]
For research use only. Not for drug, diagnostic or other use.
Fig. 1. Western Blotting
Western Blotting analysis (reducing conditions) of human Akt1 in whole cell lysate of RAJI human Burkitt lymphoma cell line.Lane 1-2:
immunostaining with polyclonal anti-Akt1 antibody