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05-1003 Sigma-Aldrich

Anti-phospho-Akt (Ser473) Antibody, clone 6F5

Anti-phospho-Akt (Ser473) Antibody, clone 6F5 detects level of phospho-Akt (Ser473) & has been published & validated for use in ELISA, WB, FC, IF, IH.

Synonym: 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

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Properties

Related Categories Antibodies, Primary Antibodies More...
format   Purified
molecular weight   60 kDa
application(s)   ELISA
  Western Blotting
  Flow Cytometry
  Immunofluorescence
  Immunohistochemistry
isotype   IgG1κ
clone   Monoclonal Antibody
  6F5
purification method   Protein G Purified
epitope   Phosphorylated Ser473
concentration   Please refer to the Certificate of Analysis for the lot-specific concentration.
host   Mouse
species reactivity   Human, Mouse, Rat, Vertebrates
NCBI accession no.   NP_001014431
UniProt accession no.   P31749
gene symbol   AKT
  C-AKT
  EC 2.7.11.1
  MGC99656
  PKB-ALPHA
  PRKBA
  RAC-ALPHA
  RAC-PK-ALPHA
  RAC
modifications   Phosphorylation
material size   100 µg
shipped in   wet ice
storage conditions   Stable for 1 year at 2-8ºC from date of receipt.

Description

Background information

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, Akt2/PKB, and Akt3/PKB)exists. They exhibit a high degree of homology, but differ slightly in the localization of their regulatory phosphorylation sites. Akt 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.

Physical form

Purified mouse monolconal IgG1κ in 0.1M Tris-Glycine (pH 7.4), 150mM NaCl with 0.05%NaN3

Application

Immunofluroescence Analysis:
EGF-treated A431 cells were fixed, permeabilized, and stained with Anti-phospho-Akt (Ser473), clone 6F5 (2 μg/mL) (red). Cy3-conjugated anti-mouse was used for detection and Phalloidin-AlexaFluor®488 (green) was used to stain actin.

Immunohistochemistry:
Anti-phospho-Akt (Ser473), clone 6F5 was diluted to 1:200, IHC-Select Detection with HRP-DAB.

Flow Cytometry:
Anti-phospho-Akt (Ser473) was used at a 1:800 dilution for this assay.

Specificity

Recognizes Akt only when phosphorylated on Ser473. This antibody does not cross-react with Akt1/PKBα when it is not phosphorylated on Ser473.

Immunogen

Peptide encompassing and including phosphorylated Ser473 of human Akt1.

Species Reactivity Notes

Human, mouse, and rat tested. Expected to react with most species as sequence is 100% homologous with most common vertebrate.

Quality Assurance

Western Blot Analysis:
0.5-1 μg/mL of this lot detected phosphorylated Akt in lysates from mouse NIH-3T3 fibroblasts treated with 100 ng/mL PDGF for 20 minutes.

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Autophagy and Mitophagy in Tumor Cell Survival and Death

Autophagy is a highly regulated process that is involved in cell growth, development, and death. In autophagy cells destroy their own cytoplasmic components in a very systematic manner and recycle th...
Chandra Mohan, Ph.D.
MilliporeSigma, Temecula, CA
Keywords: Apoptosis, Autophagy, Bacterial conjugations, Cancer, Cell proliferation, Degradations, Diseases, Immunofluorescence, Oxidations, Phosphorylations, Reductions, Separation, Ubiquitinations, Western blot

Peer-Reviewed Papers
15

References

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