Akt1, phosphorylated (Thr308) (Rac PKa, PKBa)

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 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 nhibition of apoptosis and promotion of proliferation.\n\nApplications:\nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution: \nWestern Blot: 1:1,000-1:5,000 detects phospho-Akt1/PKBa in RIPA lysates from 3T3/NIH cells stimulated with 100ng/ml PDGF for 20 minutes. Lysates from untreated (Lanes 1\nand 3) and PDGF treated (Lanes 2 and 4) 3T3/NIH cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-phospho-Akt1/PKBa (Thr308) (1:2,500) or total anti-Akt1/PKBa (1:1,000). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

A1125-18A

Size

100ul

Applications

Hosts

Rabbit

Reactivities

Hum, Mouse

Form

Supplied as a liquid in 0.2M Tris-glycine, pH 7.2, 0.15M NaCl and 0.05% sodium azide before the addition of glycerol to 30%.

P Type

Mab

Purity

Purified by Protein A affinity chromatography.

Isotype

IgG

References

1. Alessi D.R., et al., EMBO 15: 6541-6551, 1996. 2. Cross, D.A., et al., Nature 378: 785-789, 1995. 3. James, S.R., et al., Biochem. J. 315: 709-713, 1996. 4. Burgering and Coffer, Nature 376: 599-601, 1995. 5. Ackler, S., et al., Oncogene 21: 198-206, 2002. General References:1. Spieker-Polet H, et al. Proc Natl Acad Sci. 1995 Sep 26;92(20):9348-52. 2. Liguori MJ, et al. Hybridoma. 2001 Jun; 20(3):189-98. 3. G.Cano1, F. Milanezi2, D. Leitao2,3, S. Ricardo2, M.J. Brito1, F. C. Schmitt2-3 1Garcia da Orta Hospital, Almada, Portugal,2 Inst. Molec. Pathology and Immunology of Porto University, Portugal,3 Medical Faculty of Porto university, Portugal Diagn Cytopathol, 2003 Oct; 29(4): 207 -11. 4. L.K. Diaz* and N.Sneige *Department of Pathology,Northwestern University, Chicago,+ Department of Pathology, University of Texas, Huston, Adv Anat Pathol,2005; 12(1), 10-19. 5. Z. Huang1, W. Zhu2, G. Szekeres3, H. Xia1 1Spring Bioscience Corp, Fremont,CA, 2 Epitomics Inc, Burlingame,CA, , 4Histopathology Ltd, Hungary, Appl Immunohistochem Mol Morphol. 2005; 13 (1): 91-95 6. S. Rossi1, E. Orvieto1, S.Chinellato1, A. Furlanetto1, L.Laurino1, F. Facchetti2, AP Dei Tos 2 1Department of Pathology, 2Treviso, Italy; *Brescia, University School of Medicine, Brescia, Italy., Abstract presented at USCAP 2004. Modern Pathology 2004; 17 (suppl 1): 361A 7. M. Blechner, E. Ballesteros, D. Mandich, D. Stevens, R. Cartun, Hartford Hospital, Hartford, CT. Abstract presented at USCAP 2004. Modern Pathology 2004; 17 (suppl 1): 241A 8. W. Cheuk, K.O.Y. Wong, C.S.C. Wong and J.K.C. Chan, Department of Pathology, Queen Elizabeth Hospital, Hong Kong, Am J Surg Path, 2004; 28 (6): 801-807. 9. G.B. Budd, E. Tso, B. Yoder, T. Choueiri, P. Elson, S. Tarr, M. Skacel, R. Tubbs, A. Dawson, D. Hicks, Cleveland Clinic Foundation, Cleveland, OH, Abstract presented at ASCO Annual meeting, June 2004, New Orleans 10. S. M. Tarr, S. Short, K. Hansen, T. Morken, H. Xia, E. Downs-Kelly, R. R. Tubbs, D. G. Hicks, Department of Pathology and Laboratory Medicine. The Cleveland Clinic Foundation, Cleveland, Ohio. Lab Vision Corp., Fremont, Ca., Spring Bioscience Corp, Fremont ,CA, Abstract presented at Association for Molecular Pathology meeting, Los Angeles, 2004 11. A.M. Gown, T.S. Barry, P. Kandalaft, L.C. Goldstein, C.C. Tse and D.O. Treaba, Clinical Research Division , PhenoPath Laboratories and IMPRIS, Seattle, WA, Abstract presented at USCAP 2005. Modern Pathology 2005; 18, suppl.1,pag 35A 12. D.O. Treaba, A.W. Hing, L.C. Goldstein, T.S. Barry, P. Kandalaft, C.B. Gilks, T.O. Nielsen and A.M. Gown, Clinical Research Division , PhenoPath Laboratories and IMPRIS, Seattle, WA, USA Genetic Pathology Evaluation Centre, University of British Columbia, Vancouver, BC, Canada, Abstract presented at USCAP 2005. Modern Pathology 2005; 18, suppl.1,pag 53A 13. S. Rossi1, L. Laurino1, A. Furlanetto1, S.Chinellato1, E. Orvieto1, F. Canal1, F. Facchetti2, A.P. Dei Tos1 1 Depart. Pathology, Hospital of Treviso, Italy, 2 Brescia University School of Medicine, Brescia, Italy, Am J Clin Pathol, 2005, Aug;124(2):295-302

Additional Info

Recognizes phosphorylated Akt1/PKBa, Mr 60kD. Crossreactivity observed with Akt2, but not with Akt3. Species Crossreactivity: Human and mouse. Predicted to crossreact with rat, bovine and chicken based on sequence homology. 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.