Product Name: Akt1-pS473+pY474

Product Number: AB-PK514

Size:	25 µg		Price:	89.00
			$US

Target Full Name: RAC-alpha serine/threonine-protein kinase

Target Alias: Akt; Akt1; Akt1 kinase; C-Akt; Kinase Akt1; PKB; PKB-alpha; PRKBA; V-akt murine thymoma viral oncogene 1; Protein kinase B; RAC; RAC-alpha serine,threonine kinase; RAC-alpha serine/threonine kinase; RAC-PK-alpha; PKBA; hCG_96740; Akt1_NEW; Akt; MGC99656; CCDS9994; B0LPE5; B2RAM5; B3KVH4; B3KXD7; B7Z5R1; Q9BV07; ENSG00000142208

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK514

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: S473+Y474

Protein UniProt: P31749

Protein SigNET: P31749

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human Akt1 (PKBa) sequence peptide Cat. No.: PE-04ACG75

Antibody Immunogen Sequence: QF(pS)(pY)SAS(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues Q471 to S477; In Pkinase_C domain near C-terminus of the kinase

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Potency: Very strong immunoreactivity with immunogen peptide on dot blots.

Antibody Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 57-62 kDa.

Scientific Background: Akt1 (PKBA) is a protein-serine/threonine kinase of the AGC group and Akt family. It is normally inactive in the cytoplasm. When PI3K is activated by extracellular growth factors, Akt1 is recruited to the cell membrane and phosphorylated by PDK1, mTOR and other protein kinases to become active. This leads to the induction of survival, proliferation, angiogenesis and metabolism pathways. Akt1 is also involved in cell-cycle progression via its inhibitory phosphorylation of cyclin-dependent kinases and activatory phosphorylation of mTOR. This kinase is highly expressed in most tested human tissues. Orthologues of Akt are conserved in a wide range of animals, plants and unicellular eukaryotes. Phosphorylation of T72, S124,S129, S246, T308, Y315,Y326, T450, S473 and Y474 increases Akt1 phosphotransferase activity. TRAF6-induced 'K63'-linked Akt1 ubiquitination is critical for phosphorylation and activation. Phosphorylation of T34 inhibits phosphotransferase activity. Akt1 has been implicated in a wide range of human cancers, including lymphomas, breast, lung, colorectal, pancreatic and prostate cancers.

Figure 1. Epitope mapping of Akt1-pS473+pY474 antibody with similar phosphopeptides on dot blots.

Figure 2. Identification of phosphosites related to Akt1-pS473+pY474.