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Purified Mouse anti-Akt (pS473)
Purified Mouse anti-Akt (pS473)
LEFT: Western blot analysis of AKT (pS473) in mouse embryonic fibroblasts. Lysates from control (left blot) and PDGF-treated (Sigma Cat. No. P8147, right blot) NIH/3T3 cells (ATCC CRL-1658) were probed with Purified Mouse anti-Akt (pS473) monoclonal antibody at the following concentrations: 12.5 (lanes 1), 6.25 (lanes 2), and 3.125 ng/ml (lanes 3). AKT (pS473) is identified as a 60-kDa band in the treated cells. RIGHT: Western blot analysis of AKT (pS473) in  human T leukemia. Lysates from control (left panel) and Wortmannin-treated (Invitrogen, Cat. No. PHZ1301, right panel) Jurkat cells (ATCC  TIB152) were probed with Purified Mouse anti-Akt (pS473) monoclonal antibody at the following concentrations: 2.0 (lanes 1), 1.0 (lanes 2), and 0.5 µg/ml (lanes 3). The data demonstrates that the level of phosphorylation of Akt (pS473) decreases when phosphatidylinositol 3-kinase activity is inhibited.
LEFT: Western blot analysis of AKT (pS473) in mouse embryonic fibroblasts. Lysates from control (left blot) and PDGF-treated (Sigma Cat. No. P8147, right blot) NIH/3T3 cells (ATCC CRL-1658) were probed with Purified Mouse anti-Akt (pS473) monoclonal antibody at the following concentrations: 12.5 (lanes 1), 6.25 (lanes 2), and 3.125 ng/ml (lanes 3). AKT (pS473) is identified as a 60-kDa band in the treated cells. RIGHT: Western blot analysis of AKT (pS473) in  human T leukemia. Lysates from control (left panel) and Wortmannin-treated (Invitrogen, Cat. No. PHZ1301, right panel) Jurkat cells (ATCC  TIB152) were probed with Purified Mouse anti-Akt (pS473) monoclonal antibody at the following concentrations: 2.0 (lanes 1), 1.0 (lanes 2), and 0.5 µg/ml (lanes 3). The data demonstrates that the level of phosphorylation of Akt (pS473) decreases when phosphatidylinositol 3-kinase activity is inhibited.
Product Details
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BD Pharmingen™
Akt1, Akt2, Akt3, PKBα, PKBβ, PKBγ, RAC-PKα, RAC-PKβ, RAC-PKγ, STK-2
Human (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG1, κ
Phosphorylated Human Akt1 (pS473) Peptide
Western blot (Routinely Tested), Intracellular staining (flow cytometry) (Tested During Development)
60 kDa
0.5 mg/ml
AB_1645553
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. An isotype control should be used at the same concentration as the antibody of interest.
560397 Rev. 2
Antibody Details
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M89-61

Akt [also known as PKB (Protein kinase B) or RAC-PK (Related to the A and C kinases)] is a family of serine/threonine kinases that contains a pleckstrin homology (PH) domain. PH domains play important roles in signal transduction.  There are three known isoforms of Akt in mammalian cells [Akt1 (α), Akt2 (β) and Akt3 (γ)]; they are thought to be regulated similarly.  Akt is activated by insulin and growth factors by a mechanism involving phosphoinositide 3-OH kinase.  Phosphoinositide 3-OH kinases products bind to the PH domain, resulting in translocation of Akt to the plasma membrane and activation of Akt to phospho-Akt by upstream kinases.  Akt is phosphorylated within the activation loop at threonine 308 and the C-terminus at serine 473 (S473).  Phospho-Akt promotes cell survival by inhibiting apoptosis.  Specifically, phospho-Akt1 has been shown to phosphorylate Bad, a member of the Bcl-2 family that promotes cell death.  This phosphorylation results in the inactivation of the proapoptotic function of Bad.  The Akt molecule is thus considered to link extracellular survival signals (growth factors) with the apoptotic machinery (BAD).  Akt is also a key mediator of the metabolic effects of insulin.  Additionally, Akt has been referred to as an oncogene because it has increased activity in a number of tumors.

The M89-61 antibody recognizes Akt phosphorylated at S473.  This phosphorylation site is shared by all three isoforms of Akt.  The homologous phosphorylation sites in Akt2 and Akt3 are S474 and S472, respectively.

560397 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
560397 Rev.2
Citations & References
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Development References (5)

  1. Alessi DR, Andjelkovic M, Caudwell B, et al. Mechanism of activation of protein kinase B by insulin and IGF-1. EMBO J. 1996; 15(23):6541-6551. (Biology). View Reference
  2. Cantley LC, Neel BG. New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway. Proc Natl Acad Sci U S A. 1999; 96(8):4240-4245. (Biology). View Reference
  3. Datta SR, Dudek H, Tao X, et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell. 1997; 91:231-241. (Biology). View Reference
  4. Ferrigno P, Silver PA. Regulated nuclear localization of stress-responsive factors: how the nuclear trafficking of protein kinases and transcription factors contributes to cell survival. Oncogene. 1999; 18(45):6129-6134. (Biology). View Reference
  5. Kandel ES, Hay N. The regulation and activities of the multifunctional serine/threonine kinase Akt/PKB. Exp Cell Res. 1999; 253(1):210-229. (Biology). View Reference
View All (5) View Less
560397 Rev. 2

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.