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Primary Antibodies
Akt pan Antibody
Background
Akt, also known as Protein Kinase B (PKB) or RAC-α, is a 60 kDa serine/threonine kinase that plays an important role in multiple biological responses including metabolism, cell survival and growth by phosphorylating many proteins including GSK-3β, caspase 9, BAD, and the Forkhead Transcription Factor. Akt is activated by PI3K, which in turn can interact with proteins like FAK thereby linking activation to the cytoskeleton. Akt/PKB is constitutively active in many types of human cancer, and contributes to tumor progression by promoting angiogenesis and cell invasiveness. Akt is fully activated by phosphorylation of threonine 308 by PDK1 and serine 473 by PDK2.
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Description
Extracts of 3T3L1 cells were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with a 1:1000 (1) or 1:500 (2) dilution of the Akt/PKB pan antibody for two hours at room temperature in a 3% BSA-TBST buffer. After washing, the membrane was incubated with goat F(ab’)2 anti-rabbit IgG HRP-conjugate and signals were detected using the Pierce SuperSignal™ method.
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Source
Akt pan antibody was produced against a chemically synthesized phosphopeptide derived from the carboxy-terminus region of human Akt1.
Purification
Akt pan antibody was purified from rabbit serum by sequential epitope-specific chromatography. The final product is generated by affinity chromatography using a carboxy-terminal Akt peptide.
Clonality / Clone
This is a polyclonal antibody.
Host
Akt pan antibody was raised in rabbit.
Please use anti-rabbit secondary antibodies.
Specificity
Human and mouse Akt1. Rat Akt1 (94% homologous), human Akt2 (65% homologous) and human Akt3 (82% homologous) have not been tested but are expected to react.
Application
For Western blotting applications, we recommend using the antibody at a 1:1000 starting dilution.
Tested Application
WB
Buffer
Rabbit polyclonal immunoglobulin in Dulbecco’s phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol with 1.0 mg/mL BSA (IgG, protease free) as a carrier.
Storage
Store at -20˚C. We recommend a brief centrifugation before opening to settle vial contents. Then, apportion into working aliquots and store at -20˚C. For shipment or short-term storage (up to one week), 2-8˚C is sufficient.
Positive Control
- 3T3L1 extracts
Species Reactivity
H, M
Formulation
Rabbit polyclonal immunoglobulin in Dulbecco’s phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol with 1.0 mg/mL BSA (IgG, protease free) as a carrier.
Short Description
Akt pan
This product belongs to the following categories:
References- Persad, S., et al. (2001) Regulation of protein kinase B/Akt-threonine-308 phosphorylation by integrin linked kinase (ILK): Critical roles for kinase activity and amino acids arginine-211 and threonine-343. J. Biol. Chem. 276(29):27462-27469.
- Nelson, J.M. and D.W. Fry. (2001) Akt, MAPK (Erk1/2), and p38 act in concert to promote apoptosis in response to ErbB receptor family inhibition. J. Biol. Chem. 276(18):14842-14847.
- Genot, E.M., et al. (2000) The T-Cell receptor regulates Akt (Protein Kinase B) via a pathway involving Rac1 and phosphotidylinositide 3-Kinase. Mol. Cell. Biol. 20(15):5469-5478.
- Datta, S.R., et al. (1997) Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91(2):231-241.
Datasheet 0801-W
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