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.
Datasheet:
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Publication Reference
1.
Phaseolus vulgaris Exerts an Inhibitory Effect on Platelet Aggregation through AKT Dependent Way.Rodríguez-Azúa R, Quinteros EF, Olate-Briones A, Moore-Carrasco R.Prev Nutr Food Sci. 2018 Jun;23(2):102-107. doi: 10.3746/pnf.2018.23.2.102. Epub 2018 Jun 30.
2.
PKB/Akt: a key mediator of cell proliferation, survival and insulin responses?Lawlor MA, Alessi DR.J Cell Sci. 2001 Aug;114(Pt 16):2903-10.
3.
Discovery of PDK1, one of the missing links in insulin signal transduction. Colworth Medal Lecture.Alessi DR.Biochem Soc Trans. 2001 May;29(Pt 2):1-14.
4.
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.Proc Natl Acad Sci U S A. 1991 May 15;88(10):4171-5.
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 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.
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
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