AKT1 (phospho S473) monoclonal antibody, clone 17F6.B11
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Specifications
Product Description
Mouse monoclonal antibody raised against synthetic phosphopeptide of AKT1.
Immunogen
Synthetic phosphopeptide corresponding to residues surrounding S473 of human AKT1.
Host
Mouse
Reactivity
Chimpanzee, Human, Mouse, Rat
Specificity
This antibody is specific to human and mouse AKT protein phosphorylated at S473.
Form
Liquid
Isotype
IgG1, kappa
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
ELISA (1:20000)
Western Blot (1:500-1:3000)
Immunohistochemistry (20 ug/mL)
The optimal working dilution should be determined by the end user.Storage Buffer
In 20 mM KH2PO4, 150 mM NaCl, pH 7.2 (0.01% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Cell lysate)
Western blot using AKT1 (phospho S473) monoclonal antibody, clone 17F6.B11 (Cat # MAB1919) shows detection of phospho-AKT1 (indicated by arrowhead at ~56 KDa) on PDGF stimulated NIH/3T3 cell lysates (Lane 2).
No reactivity is seen for non-phosphorylated AKT1 in untreated cells (Lane 1).
Each lane contained approximately 10 ug of lysate.
All samples were loaded on to a 4-20% gradient gel for separation.
After electrophoresis, the gel was blocked with 5% BLOTTO in TBS for 90 min at RT.
The membrane was probed with the primary antibody at a 1:10,000 dilution in TBS with 0.05% Tween-20 with 1% BSA, for 1 h at 4°C. For detection HRP conjugated Gt-a-Mouse IgG was used at a 1:20,000 dilution for 1 h at 4°C with FemtoMax™ enhanced chemiluminescent reagent.
Images were captured using 2X2 binning for 10-20 sec using a BioSpectrum Imaging System (UVP Ltd.).Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Immunohistochemistry of formalin fixed and paraffin embedded using AKT1 (phospho S473) monoclonal antibody, clone 17F6.B11 (Cat # MAB1919) shows detection of Phospho-AKT1 S473 in human prostate tissue.
The antibody was used at 20 ug/mL .
The staining is much stronger than the weak basal level of phosphorylation in normal prostate tissue.
Signal was developed using Dako's Techmate streptavidin-biotin reagents.
Personal communication, Glenna Burmer, Lifespan Biosciences, Seattle, WA.Enzyme-linked Immunoabsorbent Assay
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Gene Info — AKT1
Entrez GeneID
207Protein Accession#
P31749;AAH00479Gene Name
AKT1
Gene Alias
AKT, MGC99656, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA
Gene Description
v-akt murine thymoma viral oncogene homolog 1
Gene Ontology
HyperlinkGene Summary
The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq
Other Designations
RAC-alpha serine/threonine-protein kinase|murine thymoma viral (v-akt) oncogene homolog-1|protein kinase B|rac protein kinase alpha
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Interactomes
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Pathways
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Diseases
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Publication Reference
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Phaseolus vulgaris Exerts an Inhibitory Effect on Platelet Aggregation through AKT Dependent Way.
Rodríguez-Azúa R, Quinteros EF, Olate-Briones A, Moore-Carrasco R.
Preventive Nutrition and Food Science 2018 Jun; 23(2):102.
Application:WB, Human, Platelets.
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PKB/Akt: a key mediator of cell proliferation, survival and insulin responses?
Lawlor MA, Alessi DR.
Journal of Cell Science 2001 Aug; 114(Pt 16):2903.
Application:IHC, WB-Tr, Human, Cancers, Mammalian cells.
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Discovery of PDK1, one of the missing links in insulin signal transduction. Colworth Medal Lecture.
Alessi DR.
Biochemical Society Transactions 2001 May; 29(Pt 2):1.
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Molecular cloning and identification of a serine/threonine protein kinase of the second-messenger subfamily.
Jones PF, Jakubowicz T, Pitossi FJ, Maurer F, Hemmings BA.
PNAS 1991 May; 88(10):4171.
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Phaseolus vulgaris Exerts an Inhibitory Effect on Platelet Aggregation through AKT Dependent Way.
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