AKT2 polyclonal antibody
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More Files
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Western Blot (Tissue lysate)
Immunoblotting of AKT2 polyclonal antibody (Cat # PAB9933) to AKT2 was used at a 1 : 1,000 dilution to detect AKT2 by Western blot in lysates from mouse tissues.
The antibody is shown to react with AKT2 in mouse liver, skeletal muscle and fat using standard immunoblotting methods.
Western Blot (Cell lysate)
Immunoblotting of AKT2 polyclonal antibody (Cat # PAB9933) detects both unphosphorylated and phosphorylated AKT2.
As before approximately 20 mg/lane of crude HEK293 lysate was loaded for SDS-PAGE, separated and then transferred to nitrocellulose.
The right lane contains lysate pretreated for 15 min with 25 &mciro;M LY294002 which affects the phosphorylation of endogenous AKT2, but has no effect on phosphorylation of the myristoylated construct.
This antibody clearly detects both the phosphorylated (top arrow) and the non-phosphorylated (bottom arrow) forms of endogenous AKT2.
Western Blot (Transfected lysate)
Immunoblotting of AKT2 polyclonal antibody (Cat # PAB9933) to AKT2 wasused at a 1 : 1,000 dilution to detect AKT2 by Western blot.
A lysate from a stable HEK293 cell line expressing an inducible, myristoylated form of AKT2 (MyrAKT2-ER) was loaded for SDS-PAGE, separated and then transferred to nitrocellulose.
The blot was reacted with primary antibody for 1h at room temperature.
Inresponse to 4OHT (tamoxifen), the AKT2 is recruited to the plasma membrane via its myristoylation sequence and becomes phosphorylated and activated.
The blot (right panel) shows 0 m, 15m, and 30 m of 4OHT treatment.
This treatment has no effect on endogenous AKT2, but causes a band shift upwards in the MyrAKT2.
Endogenous AKT2 runs at about 60 KDa whereas the myristoylated construct runs at around 110 KDa.
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptide of AKT2.
Immunogen
A synthetic peptide (conjugated with KLH) corresponding to amino acids 455-468 of human AKT2.
Sequence
RYDSLGLLELDQRT
Host
Rabbit
Reactivity
Human, Mouse, Rat
Specificity
This antibody detects both unphosphorylated and phosphorylated forms of the protein.
Form
Liquid
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
ELISA (1:4000-1:16000)
Western Blot (1:1000)
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 (Tissue lysate)
Immunoblotting of AKT2 polyclonal antibody (Cat # PAB9933) to AKT2 was used at a 1 : 1,000 dilution to detect AKT2 by Western blot in lysates from mouse tissues.
The antibody is shown to react with AKT2 in mouse liver, skeletal muscle and fat using standard immunoblotting methods.Western Blot (Cell lysate)
Immunoblotting of AKT2 polyclonal antibody (Cat # PAB9933) detects both unphosphorylated and phosphorylated AKT2.
As before approximately 20 mg/lane of crude HEK293 lysate was loaded for SDS-PAGE, separated and then transferred to nitrocellulose.
The right lane contains lysate pretreated for 15 min with 25 &mciro;M LY294002 which affects the phosphorylation of endogenous AKT2, but has no effect on phosphorylation of the myristoylated construct.
This antibody clearly detects both the phosphorylated (top arrow) and the non-phosphorylated (bottom arrow) forms of endogenous AKT2.Western Blot (Transfected lysate)
Immunoblotting of AKT2 polyclonal antibody (Cat # PAB9933) to AKT2 wasused at a 1 : 1,000 dilution to detect AKT2 by Western blot.
A lysate from a stable HEK293 cell line expressing an inducible, myristoylated form of AKT2 (MyrAKT2-ER) was loaded for SDS-PAGE, separated and then transferred to nitrocellulose.
The blot was reacted with primary antibody for 1h at room temperature.
Inresponse to 4OHT (tamoxifen), the AKT2 is recruited to the plasma membrane via its myristoylation sequence and becomes phosphorylated and activated.
The blot (right panel) shows 0 m, 15m, and 30 m of 4OHT treatment.
This treatment has no effect on endogenous AKT2, but causes a band shift upwards in the MyrAKT2.
Endogenous AKT2 runs at about 60 KDa whereas the myristoylated construct runs at around 110 KDa.Enzyme-linked Immunoabsorbent Assay
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Gene Info — AKT2
Entrez GeneID
208Gene Name
AKT2
Gene Alias
PKBB, PKBBETA, PRKBB, RAC-BETA
Gene Description
v-akt murine thymoma viral oncogene homolog 2
Gene Ontology
HyperlinkGene Summary
This gene is a putative oncogene encoding a protein belonging to a subfamily of serine/threonine kinases containing SH2-like (Src homology 2-like) domains. The gene was shown to be amplified and overexpressed in 2 of 8 ovarian carcinoma cell lines and 2 of 15 primary ovarian tumors. Overexpression contributes to the malignant phenotype of a subset of human ductal pancreatic cancers. The encoded protein is a general protein kinase capable of phophorylating several known proteins. [provided by RefSeq
Other Designations
Murine thymoma viral (v-akt) homolog-2|rac protein kinase beta
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Interactome
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Pathway
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Disease
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Publication Reference
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Dwarfism, impaired skin development, skeletal muscle atrophy, delayed bone development, and impeded adipogenesis in mice lacking Akt1 and Akt2.
Peng XD, Xu PZ, Chen ML, Hahn-Windgassen A, Skeen J, Jacobs J, Sundararajan D, Chen WS, Crawford SE, Coleman KG, Hay N.
Genes & Development 2003 Jun; 17(11):1352.
Application：WB, Mouse, Skeletal muscles.
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Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta).
Cho H, Mu J, Kim JK, Thorvaldsen JL, Chu Q, Crenshaw EB 3rd, Kaestner KH, Bartolomei MS, Shulman GI, Birnbaum MJ.
Science 2001 Jun; 292(5522):1728.
Application：WB-Ti, Mouse, Liver, Skeletal muscle, Adipocytes.
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Amplification of AKT2 in human pancreatic cells and inhibition of AKT2 expression and tumorigenicity by antisense RNA.
Cheng JQ, Ruggeri B, Klein WM, Sonoda G, Altomare DA, Watson DK, Testa JR.
PNAS 1996 Apr; 93(8):3636.
Application：WB-Ce, WB-Tr, Human, ASPC-1, CD-11, Colo 357, HeLa, PAN1, PANC-1, T3M4 cells.
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Dwarfism, impaired skin development, skeletal muscle atrophy, delayed bone development, and impeded adipogenesis in mice lacking Akt1 and Akt2.
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