Product Name: PRKACA-pT196+pT198

Product Number: AB-PK782

Size:	25 µg		Price:	89.00
			$US

Target Full Name: cAMP-dependent protein kinase catalytic subunit alpha

Target Alias: Alpha-catalytic subunit; cAMP-dependent protein kinase; cAMP-dependent protein kinase, alpha-catalytic subunit; cAPKa; KAPCA; PKA C-alpha; PKA-alpha; PKACA; Protein kinase, cAMP-dependent, catalytic, alpha; MGC102831; MGC48865; CCDS12304.1; P17612; ENSG00000072062

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK782

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: T196+T198

Protein UniProt: P17612

Protein SigNET: P17612

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human PRKACA sequence peptide Cat. No.: PE-04AOQ85

Antibody Immunogen Sequence: KGR(pT)W(pT)LCG(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues K193 to G201; In the protein kinase catalytic domain activation T loop region between subdomains VII and VIII.

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.

Antibody Concentration: 1 mg/ml

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 38-42 kDa.

Scientific Background: PKACa (PRKACA) is a protein-serine/threonine kinase of the AGC group and PKA family. It serves as the alpha catalytic subunit of the cAMP-dependent protein kinase holoenzyme PKA, and it phosphorylates and regulates a very large number of cytoplasmic and nuclear substrate proteins. Phosphorylates hundreds of substrates in the cytoplasm and the nucleus. These include CDC25B, ABL1, NFKB1, CLDN3, PSMC5/RPT6, PJA2, RYR2, RORA, TRPC1 and VASP. It is activated by binding of two cAMP molecules to each of the two associated regulatory subunits in the PKA holoenzyme. Binding of cAMP induces dissociation of the two active catalytic subunits. Phosphorylation of T198 increases phosphotransferase activity. Phosphorylation of S338 plays a key role in stabilizing PKACa. PKACa may be an oncoprotein (OP), although this kinase has also been implicated in inhibiting normal cell proliferation. Gain-of-function mutations in the PRKACA catalytic domain are associated with elevated catalytic activity of the kinase, and have been observed in several cancer types. In particular, a L206R subsitution mutation is observed in cancer and in Cushing's syndrome patients. The gain-of-function mutation is predicted to result in the interruption of the interaction between the catalytic and regulatory subunits of protein kinase A, resulting in a constitutively active PKA. Gain-of-function mutations in the PRKACA gene that lead to consitutive activity of the PKA holoenzyme have been proposed to play an important role in the development of several types of cancer.

Figure 1. Identification of phosphosites related to PRKACA-pT196+pT198.