V-Akt Murine Thymoma Viral Oncogene Homolog 1/2/3 (AKT1/AKT2/AKT3) (C-Term) antibody

Antigen: V-Akt Murine Thymoma Viral Oncogene Homolog 1/2/3 (AKT1/AKT2/AKT3)

Binding Site: C-Term

Clonality: Polyclonal

Host: Rabbit

Reactivity: Chicken

Conjugate: Un-conjugated

Application: Immunofluorescence (IF), Immunoprecipitation (IP), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB), Immunohistochemistry (IHC), ELISA

Catalog no.: ABIN117897

Additional Information

Alternative name: AKT1/AKT2/AKT3

UniProt: P31749

Immunogen: Synthetic peptide corresponding to the C-terminus (460-480) of Human, Rat and Mouseand chicken AKT proteins conjugated to KLH using maleimide. A residue of cysteine wasadded to the amino terminal end to facilitate coupling. AA Sequence: C-R-PH-F-P-Q-F-S-Y-S-A-S-G-T-A

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

Description: AKT is a component of the PI-3 kinase pathway and is activated by phosphorylation at Ser473 and Thr 308. AKT is a cytoplasmic protein also known as Protein Kinase B (PKB) andrac (related to A and C kinases). AKT is a key regulator of many signal transductionpathways. AKT Exhibits tight control over cell proliferation and cell viability. Overexpression or inappropriate activation of AKT is noted in many types of cancer. AKTmediates many of the downstream events of PI 3-kinase (a lipid kinase activated by growthfactors, cytokines and insulin). PI 3-kinase recruits AKT to the membrane, where it isactivated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from themembrane and phosphorylates targets in the cytoplasm and the cell nucleus. AKT has twomain roles: (i) inhibition of apoptosis, (ii) promotion of proliferation.

Characteristics: Synonyms: Akt-1, RAC-PK-alpha, Protein kinase B, C-AKT, RAC-alpha serine/threonine-protein kinase

Specificity: This antiserum reacts with the AKT. Species Reactivity: Tested: Human. Expected from sequence similarity: Rat, Mouse and Chicken.

Application Details

Application Notes: ELISA: 1/2,000-1/10,000. Immunoprecipitation. Transfected/Primary cell culture. Western Blot (1/500-1/2,000): Detect a single band of the expected apparent molecularweight. Immunofluorescence using paraformaldehyde-fixed primary cardiomyocyte cultures(1/100-1/1,000). Immunohistochemistry on Formalin-Fixed Paraffin-Embedded Sections (1/500-1/2,000). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Concentration: 85 mg/ml (by refractometry)

Purification: Serum

Buffer: 0.02M Potassium Phosphate, 0.15M Sodium Chloride, pH 7.2 containing0.09% Sodium Azide

Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Shelf life: one year from despatch.

Restrictions: For Research Use only

Images

Immunofluorescence microscopy: Rabbit anti-AKT was used at a 1:80 dilution to stain cultured neonatal rat cardiomyocytes that express a nuclear-targeted AKT construct. Anti-AKT staining appears green. Actin filaments are labeled red using a Texas-red™ conjugated phalloidin.

Immunoblotting: Rabbit anti-AKT was used at a 1:500 dilution to detect AKT by Western blot. Approximately 20 ug of an NIH/3T3 whole cell lysate was loaded on a 10% NuPage SDS-PAGE gel using MOPS buffer. After washing, a 1:10,000 dilution of HRP conjugated Gt-a-Rabbit IgG (R1454HRP) preceded color development using Pierce Chemical's SuperSignal™ substrate.

Immunohistochemistry. Rabbit anti-AKT was used at a 1:1,000 dilution to detect AKT by immunohistochemistry..,.In panel (A) normal colon tissue is stained. Panel (B) shows tumor tissue. Tissue was formalin-fixed and paraffin embedded.

Publications

Lawlor, Alessi: "PKB/Akt: a key mediator of cell proliferation, survival and insulin responses?" in: Journal of cell science, Vol. 114, Issue Pt 16, pp. 2903-10, 2001 (PubMed).