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Primary Antibodies  >  Phospho Specific Antibodies
 

Akt phospho S473 Antibody

Mouse Monoclonal 17F6.B11 IgG1 kappa
Code

200-301-B19
Size

1 mg
Price

$1,989.00
Availability

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Immunofluorescence Microscopy of Mouse Anti-AKTpS473 antibody using STED nanoscopy to evaluate AKT activation and migration.  
Tissue:  A431 cells.  
Antigen retrieval:  Panel A: serum starved,unstimulated cells.  Panel B: serum starved, EGF stimulated for 15 mins.
A massive increase in AKT-pS473 activation, as measured by intensity signal, peaked at 15 minutes and was associated with depolymerized tubulin.
Staining:  Panel A shows STED data (AKT-pS473, red channel) collected simultaneously with confocal signal (a-tubulin, green channel).  Upon stimulation of cells with EGF, a rapid activation of AKT is observed (Panel B) along with a coincident change in the tubulin organization (yellow signal), as well as an extensive cell shape-change (cell membrane folding) and accumulation of AKTpS473 at the cell periphery.
Western Blot of Mouse Anti-AKTpS473 antibody.  
Lane 1:  A431 cells.  
Lane 2:  A431 cells stimulated for 15 min with EGF.  
Load:  35 µg per lane.
Primary antibody:  AKTpS473 antibody at 1:400 for overnight at 4°C.
Secondary antibody:  DyLight™649 Conjugated Anti-AKT pS473 Monoclonal Antibody p/n 200-343-268 at 1:10,000 for 45 min at RT.
Block:  Blocking Buffer for Fluorescent Western Blotting p/n MB-070 overnight at 4°C.
Predicted/Observed size:  56kDa. 
Other band(s): none.
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Synonyms: RAC-PK-alpha, Protein kinase B, PKB, C-AKT, RAC-alpha serine/threonine-protein kinase, Proto-oncogene c-Akt, AKT1, AKT 1, AKT-1

Akt phospho S473 Antibody Properties

Description
Anti-AKT pS473 (MOUSE) Monoclonal Antibody - 200-301-B19
Host
Mouse
Target Species
Human
Known Cross Reactivity
human, mouse, rat, monkey
Clonality
Monoclonal 17F6.B11 IgG1 kappa
Application
ELISA : 1:20,000
IF Microscopy : 1:500 - 1:3,000
FlowCytometry : User Optimized
Western Blot : 1:500 - 1:3,000
Immunochemistry: 20 µg/ml
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG1
Label
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
AKT1
Download CofA
Lot#  

Akt phospho S473 Antibody Description

Background
AKT is a component of the PI-3 kinase pathway and is activated by phosphorylation at Ser 473 and Thr 308. AKT is a cytoplasmic protein also known as AKT1, Protein Kinase B (PKB) and rac (related to A and C kinases). AKT is a key regulator of many signal transduction pathways. AKT Exhibits tight control over cell proliferation and cell viability. Overexpression or inappropriate activation of AKT is noted in many types of cancer. AKT mediates many of the downstream events of PI 3-kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI 3-kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus. AKT has two main roles: (i) inhibition of apoptosis; (ii) promotion of proliferation. Anti-AKT pS473 (MOUSE) Monoclonal Antibody is ideal for investigators involved in Cell Signaling, Cancer, Neuroscience, Signal Transduction research.
Immunogen
This monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S473 of human AKT1 protein.
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
This monoclonal antibody is suitable for ELISA, immunohistochemistry, immunoprecipitation, immunofluorescent microscopy, and western blotting.  Expect a band approximately 56 kDa in size corresponding to phosphorylated AKT protein by western blotting in the appropriate cell lysate or extract.  This phospho-specific monoclonal antibody reacts with human and mouse AKT pS473 and shows minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide.  Specific conditions for reactivity should be optimized by the end user.  For immunohistochemistry use formalin-fixed paraffin-embedded sections.    No pre-treatment of sample is required. Cell Signaling, Cancer, Neuroscience, Signal Transduction research.
Purity/Specifity
This product was purified from concentrated tissue culture supernate by Protein A chromatography. This antibody is specific for human and mouse AKT protein phosphorylated at S473.  A BLAST analysis was used to suggest cross-reactivity with AKT pS473 from human, mouse, rat and chimpanzee sources based on 100% homology with the immunizing sequence.  Cross-reactivity with AKT from other sources has not been determined.  Cross-reactivity with AKT2 and AKT3 has not been determined.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Lawlor, M. A. and Alessi, D.R. (2001). PKB/AKT: a key mediator of cell proliferation, survival and insulin responses. J. Cell Science 114:2903-2910. Alessi, D. R. (2001). Discovery of PDK1, one of the missing links in insulin signal transduction. Biochem. Soc. Trans. 29,1 -14. Jones,P.F., Jakubowicz,T., Pitossi,F.J., Maurer,F. and Hemmings,B.A. (1991) Molecular cloning and identification of a serine/threonine protein kinase of the second-messenger subfamily. Proc. Natl. Acad. Sci. U.S.A. 88 (10), 4171-4175.
Specific Reference
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Product Type
Proteins and Peptides;
Catalog Number
000-000-401

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, monkey
Catalog Number
200-301-268

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, FC
Reacts With
human, mouse, rat, monkey
Catalog Number
200-301-269

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, FC
Reacts With
human, mouse, rat, chimpanzee
Catalog Number
200-301-401

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry