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Anti-phospho-Akt1/PKBα (Thr308), clone NL50
Recommended Replacement for: 05-802
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PRODUCT FAMILY INFORMATION
Phospho-Akt Antibodies, Kits & Assays
Millipore’s phosphoAkt antibodies have been well validated and published. See below or a comprehensive list of our phosphoAkt products, based on the expertise of Upstate & Chemicon.
EMD Millipore’s phosphoAkt antibodies are based on the expertise of Upstate & Chemicon. Our phosphoAkt antibodies have been well validated and published. See the ordering tab for a comprehensive list of our phosphoAkt products and direct links to their technical pages for data, references and related products.
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Species Reactivity Key Applications Host Format Antibody Type
H, M, B, Ch, R WB Rabbit null Monoclonal Antibody
Description:
Anti-phospho-Akt1/PKBα (Thr308) Antibody, clone NL50 | 04-802
Replaces:
05-802
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Trade Name:
Upstate (Millipore)
Specificity:
Recognizes phosphorylated Akt1/PKBα, Mr 60 kDa. Cross-reactivity observed with Akt2, but not with Akt3.
Molecular Weight:
~60 kDa
Epitope:
pThr308
Immunogen:
KLH-conjugated, synthetic peptide containing the sequence ..MK[pT]FC.. in which pT corresponds to phospho-threonine at residue 308 of mouse Akt. The Akt1 sequence is identical to human, rat, bovine and chicken Akt1. Human and mouse Akt2 shares 11/12
amino acids and rat Akt3 shares 10/12 amino acids with the peptide immunogen sequence.
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Modifications:
Phosphorylation
Clone:
NL50
Isotype:
IgG
Background Information:
Akt is the major known effector of the PI3 Kinase pathway. Generation of PIP3 results in the activation of PDK1, which phosphorylates Akt on Thr308, and another kinase complex, thought to be the mTORC2 complex (or possibly an intermediate kinase linking the two) that phosphorylates Akt on Ser473. These phosphorylations additively activate Akt Ser/Thr kinase activity, and the use of phosphorylation state-specific antibodies directed against either of these sites can imply Akt activation. Activation of Akt can be measured directly by immunoprecipitation followed by phosphorylation of a known substrate with radiolabeled ATP. Akt has been shown to phosphorylate over 70 substrates including TSC1&2 in the mTOR pathway, Bad on Ser136, GSK3, which is inactivated by this phosphorylation, the FoxO family of transcription factors, PRAS40, AS160.
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Species Reactivity:
Human
Mouse
Bovine
Chicken
Rat
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Species Reactivity Note:
Human and mouse. Predicted to cross-react with rat, bovine and chicken based on sequence homology.
Application Notes:
Immunoblot Analysis: A 1:500-1:2,000 dilution of this lot detected phospho-Akt1/PKBα in RIPA lysates from 3T3/NIH cells stimulated with 100ng/ml PDGF for 20 minutes (Figure A).
Beadlyte® Specificity Assay: 1:1,000-1:10,000 dilutions of this lot were incubated with phosphorylated and unphosphorylated recombinant Akt proteins conjugated to Luminex microspheres. Cross-reactivity to active Akt2 was detected (Figure B).
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Control:
PDGF treated 3T3/NIH cells
Quality Assurance:
Evaluated by western blot on RIPA lysates from 3T3/NIH cells stimulated with 100 ng/mL PDGF for 20 minutes.
Western Blotting Analysis: A 1:500 dilution of this antibody detected phospho-Akt1/PKBα in RIPA lysates from 3T3/NIH cells stimulated with 100 ng/mL PDGF for 20 minutes.
Presentation:
Cultured supernatant containing 0.05% sodium azide.
Storage Conditions |