Product Name: PRK1-pT774
Product Number: AB-PK781
Size: 25 µg      Price:89.00
      $US
Target Full Name: Protein kinase C-related protein-serine kinase 1; Protein kinase N1

Target Alias: PAK-1; PKN; PRK1; PRKCL1; Protease-activated kinase 1; Protein kinase C-like 1; Protein kinase C-like PKN; Protein-kinase C-related kinase 1

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK781

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: T774

Protein UniProt: Q16512

Protein SigNET: Q16512

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human PRK1 (PKN1) sequence peptide Cat. No.: PE-04ADX99

Antibody Immunogen Sequence: RTS(pT)FCG(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues R771 to G777; In protein kinase catalytic domain activation T-loop 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: 0.75 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: Medium 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 125-130 kDa.

Antibody Specificity: Very high

Related Product 1: PRK1-pT774 blocking peptide

Scientific Background: PRK1 (PKN1) is a protein-serine/threonine kinase of the AGC group and PKN family. It is associated with many functions including regulation of actin cytoskeleton, cell migration, and tumour cell invastion. Its phosphotransferase activity is stimulated upon binding to Rho proteins (RHOA, RHOB and RAC1). It is activated by lipids, particularly cardiolipin and to a lesser extent by other acidic phospholipids. It is also activated by caspase-3 (CASP3) cleavage during apoptosis., as well as in vitro by limited proteolysis by trypsin. Phosphorylationof PRK1 at T774 (activation loop of the kinase domain) and S916 (turn motif) increases phosphotransferase activity. Insertional mutagenesis studies in mice support a role for this protein kinase in mouse cancer oncogenesis. Stimulation of the PRK1 signalling cascade results in androgen receptor superactivation in human prostate cancer cells. PRK1 also represses Wnt signalling in human melanoma cells.
Figure 1. Epitope mapping of PRK1-pT774 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to PRK1-pT774.