V-Akt Murine Thymoma Viral Oncogene Homolog 2 (AKT2) antibody

Antigen: V-Akt Murine Thymoma Viral Oncogene Homolog 2 (AKT2)

Synonyms: PKBB, PRKBB, PKBBETA, RAC-BETA, AKT2, cb945, prkbb, v-akt2, pkbbeta, rac-beta

Clonality: Polyclonal

Host: Rabbit

Reactivity: Human, Mouse (Murine), Rat (Rattus)

Conjugate: Un-conjugated

Application: Western Blotting (WB)

Catalog no.: ABIN117900

Additional Information

Alternative name: AKT2 / PKB beta

UniProt: P31751

Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeatedimmunizations with a synthetic peptide of human AKT2 conjugated to Keyhole LimpetHemocyanin (KLH). AA Sequence: Synthetic peptide R-Y-D-S-L-G-L-L-E-L-D-Q-R-T (corresponding to aa 455-468of 481)

Cross-Reactivity: Human, Mouse (Murine), Rat (Rattus)

Description: AKT2 is also known as V-AKT Murine Thymoma Viral Oncogene Homolog 2 antibody, Racprotein kinase beta antibody, PKB beta antibody or PRKBB antibody. AKT2 is an isoform ofthe phosphoinositidedependent serine-threonine protein kinase AKT and is enriched ininsulin-responsive tissues and has been implicated in the metabolic actions of thehormone. AKT2 is a putative oncogene encoding a protein belonging to a subfamily ofserine/threonine kinases containing SH2-like (Src homology 2- like) domains. Furthermore,AKT2 was shown to be amplified and overexpressed in 2 of 8 ovarian carcinoma cell linesand 2 of 15 primary ovarian tumors. Overexpression of AKT2 contributes to the malignantphenotype of a subset of human ductal pancreatic cancers. AKT2 is a general proteinkinase capable of phophorylating several known proteins. AKT2 mediates many of thedownstream events of PI 3-kinase (a lipid kinase activated by growth factors, cytokinesand insulin). PI 3-kinase recruits AKT2 to the membrane, where it is activated by PDK1phosphorylation. Once phosphorylated, AKT2 dissociates from the membrane andphosphorylates targets in the cytoplasm and the cell nucleus. AKT2 has two main roles: (i)inhibition of apoptosis, (ii) promotion of proliferation.

Characteristics: Synonyms: RAC-PK-beta, Protein kinase Akt-2, Protein kinase B beta, RAC-betaserine/threonine-protein kinase

Application Details

Application Notes: This antibody was tested by ELISA and Immunoblotting and was found to be reactive withboth unphosphorylated and phosphorylated AKT2 in a lysate of HEK293 cells. Although nottested, this antibody is likely functional in Immunohistochemistry andImmunoprecipitation. Recommended Dilution(s): This product has been assayed by immunoblot against aHEK293 cell lysate and is reactive at a 1: 1,000 dilution showing a band at approximately 60

Concentration: 1.0 mg/ml

Purification: Affinity Purified

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 , with 0.01%Sodium Azide as preservative.

Storage: Store vial at -20°C prior to opening. Centrifuge product if not completely clear after standing atroom temperature. For extended storage aliquot contents and freeze at -20°C or below. Dilute only prior to immediate use. Avoid repeated freezing and thawing. Shelf life: one year from despatch.

Research Area: Signaling

Restrictions: For Research Use only

Images

Immunoblotting. Affinity Purified antibody to AKT2 was used 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 SDSPAGE, separated and then transferred to nitrocellulose. The blot was reacted with primary antibody for 1h at room temperature. In response 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, 15 m, and 30 m of 4OHT treatment. This treatment has no effect onendogenous Akt2, but causes a band shift upwards in the MyrAkt2. Endogenous Akt2 runs at about 60kDa whereas the myristoylated construct runs at around 110kDa.

Immunoblotting. Antibody to AKT2 detects both unphosphorylated and phosphorylated AKT2. As before approximately 20mg/lane of crude HEK293 lysate was loaded for SDS-PAGE, separated and then transferred to nitrocellulose. The right lane contains lysate pretreated for 15min with 25uM 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.

Publications

Cheng, Godwin, Bellacosa et al.: "AKT2, a putative oncogene encoding a member of a subfamily of protein-serine/threonine kinases, is amplified in human ovarian carcinomas." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, Issue 19, pp. 9267-71, 1992 (PubMed).

Staal: "Molecular cloning of the akt oncogene and its human homologues AKT1 and AKT2: amplification of AKT1 in a primary human gastric adenocarcinoma." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 84, Issue 14, pp. 5034-7, 1987 (PubMed).

Cheng, Ruggeri, Klein et al.: "Amplification of AKT2 in human pancreatic cells and inhibition of AKT2 expression and tumorigenicity by antisense RNA." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, Issue 8, pp. 3636-41, 1996 (PubMed).

Altomare, Kozak, Sonoda et al.: "Chromosome mapping of the mouse Akt2 gene and Akt2 pseudogene." in: Cytogenetics and cell genetics, Vol. 74, Issue 4, pp. 248-51, 1997 (PubMed).

Cho, Mu, Kim et al.: "Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta)." in: Science (New York, N.Y.), Vol. 292, Issue 5522, pp. 1728-31, 2001 (PubMed).

Peng, Xu, Chen et al.: "Dwarfism, impaired skin development, skeletal muscle atrophy, delayed bone development, and impeded adipogenesis in mice lacking Akt1 and Akt2." in: Genes & development, Vol. 17, Issue 11, pp. 1352-65, 2003 (PubMed).