Technical Data
A1124-90A
Akt, phosphorylated (Ser473) (Protein kinase B, RAC-alpha serine/threonine-protein kinase, murine thymoma viral (v-akt) oncogene homolog-1, rac protein kinase alpha, v-akt murine thymoma viral oncogene homolog 1)
Description:
Akt/PKB is a Ser/Thr kinase and a major known effecter of the PI3 Kinase pathway. It is involved in multiple signaling pathways that relate to many biological processes including glucose metabolism, cell survival/apoptosis, cell cycle control, angiogenesis, differentiation and cell growth and proliferation. In mammals three isoforms of Akt (Akt1/PKB alpha, Akt2/PKB beta, and Akt3/PKB gamma) exist. They exhibit a high degree of homology, but differ slightly in the localization of their regulatory phosphorylation sites. Akt1 is the predominant isoform that is in most tissues and is thought to have a dominant role in growth, survival, embryonic development and adipocyte differentiation. Akt2 is correlated with the regulation of glucose homeostasis and is the predominant isoform expressed in insulin- responsive tissues. Akt3 is abundant in brain tissue. Each Akt isoform is composed of three functionally distinct regions: an N-terminal Pleckstrin Homology (PH) domain that provides a lipid-binding module to direct Akt to PIP3 at the cell membrane that is necessary for its activation, a central catalytic domain containing Thr308, and a C-terminal hydrophobic motif containing Ser473. The activation of Akt is dependent on a dual regulatory mechanism that requires both its translocation to the plasma membrane and dual phosphorylation on Thr308 and Ser473 by PDK1 and the TORC2 complex, respectively.

Applications:
Suitable for use in Immunofluorescence, Flow Cytometry, ELISA, Western Blot and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Immunofluorescence: 2ug/ml
Flow Cytometry: 1:800
Western Blot: 0.5-1ug/ml detected phosphorylated AKT in lysates from mouse NIH-3T3 fibroblasts treated with 100ng/ml PDGF for 20 minutes.
Immunohistochemistry: 1:200; Pretreatment with citrate, pH 6 antigen retrieval.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgG1,k9i145Affinity Purified
SizeStorageShippingSourceHost
100ug-20CBlue IceHumanMouse
Concentration:
~1mg/ml
Immunogen:
Peptide encompassing and including phosphorylated Ser473 of human Akt1.
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 150mM sodium chloride, 0.05% sodium azide.
Specificity:
Recognizes human Akt at ~60kD only when phosphorylated on Ser473. Does not crossreact with Akt1/PKBalpha when it is not phosphorylated on Ser473. Species Crossreactivity: mouse and rat. Species sequence homology: Most vertebrates.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Cohen, P, et al. (1997) FEBS Lett., 410: 3-10 (1997). 2. James, S R, et al. (1996) Biochem. J., 315 ( Pt 3): 709-13 (1996). 3. Alessi, D R, et al. (1997) Curr. Biol., 7: 776-89 (1997). 4. Cross, D A, et al. (1995) Nature, 378: 785-9 (1995). 5. Alessi, D R, et al. (1998) Curr. Biol., 8: 69-81 (1998)