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Purified Mouse Anti-Akt (pS472/pS473)
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Purified Mouse Anti-Akt (pS472/pS473)
Western blot analysis for Akt (pS472/pS473). Serum-starved NIH/3T3 cells (Mouse embryo fibroblast cells; ATCC CRL-1658) were either left untreated (lanes 1-3) or stimulated with 10 ng/ml PDGF for 6 minutes (lanes 4-6). Blots were probed with the mouse anti-Akt (pS472/pS473) antibody at concentrations of 2 µg/mL (lanes 1, 4), 1 µg/mL (lanes 2, 5), and 0.5 µg/mL (lanes 3, 6). Note that the mouse anti-Akt (pS472/pS473) antibody recognized Akt in PDGF-treated cells (lanes 4-6), but not in untreated cells (lanes 1-3). Akt is identifiable as a band of ~60 kDa.
Western blot analysis for Akt (pS472/pS473). Serum-starved NIH/3T3 cells (Mouse embryo fibroblast cells; ATCC CRL-1658) were either left untreated (lanes 1-3) or stimulated with 10 ng/ml PDGF for 6 minutes (lanes 4-6). Blots were probed with the mouse anti-Akt (pS472/pS473) antibody at concentrations of 2 µg/mL (lanes 1, 4), 1 µg/mL (lanes 2, 5), and 0.5 µg/mL (lanes 3, 6). Note that the mouse anti-Akt (pS472/pS473) antibody recognized Akt in PDGF-treated cells (lanes 4-6), but not in untreated cells (lanes 1-3). Akt is identifiable as a band of ~60 kDa.
Product Details
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BD Pharmingen™
Akt1, Akt2, Akt3, PKBα, PKBβ, PKBγ, RAC-PKα, RAC-PKβ, RAC-PKγ, STK-2
Mouse (QC Testing), Human,Rat (Tested in Development)
Mouse IgG1
Phosphorylated Human Akt1 (pS473) Peptide
Western blot (Routinely Tested), Immunoprecipitation (Tested During Development)
0.5 mg/ml
AB_393864
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.

Recommended Assay Procedures

Western blot:  Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. 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.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
550747 Rev. 1
Antibody Details
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104A282

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 pleckstrin homology 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. 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 104A282 antibody recognizes Akt1 phosphorylated at S473 and Akt3 phosphorylated at S472. This antibody may also recognize Akt2 when phosphorylated at S474.

550747 Rev. 1
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
550747 Rev.1
Citations & References
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Development References (4)

  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. 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
  4. 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 (4) View Less
550747 Rev. 1

Please refer to Support Documents for Quality Certificates


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.