Product Name: PAK4-pS474
Product Number: AB-PK752
| Size: | 25 µg | Price: | 89.00 | |
| $US |
Target Full Name: p21-activated kinase 4
Target Alias: P21-activated kinase 4; CCDS12528.1; ENSG00000130669
Product Type Specific: Protein kinase phosphosite-specific antibody
Antibody Code: PK752
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S474
Protein UniProt: O96013
Protein SigNET: O96013
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human PAK4 sequence peptide Cat. No.: PE-04AED99
Target Alias: P21-activated kinase 4; CCDS12528.1; ENSG00000130669
Product Type Specific: Protein kinase phosphosite-specific antibody
Antibody Code: PK752
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S474
Protein UniProt: O96013
Protein SigNET: O96013
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human PAK4 sequence peptide Cat. No.: PE-04AED99
Antibody Immunogen Sequence: RRK(pS)LVG(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues R471 to G477; In the protein kinase catalytic domain activation T loop region between subdomains VII and VIII.
Antibody Immunogen Description: Corresponds to amino acid residues R471 to G477; 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 Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.5 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: Weak immunoreactivity of a target-sized protein by Western blotting in HeLa cells. Strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: Human
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: Weak immunoreactivity of a target-sized protein by Western blotting in HeLa cells. 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 67-74 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: Almost no significant cross-reactivities detected in HepG2 and HeLa cells, except 27 kDa protein in HeLa; in HepG2 cells, insulin increases 27 kDa protein.
Related Product 1: PAK4-pS474 blocking peptide
Related Product 2: PAK4Subtide - PAK4 protein kinase substrate peptide
Antibody Specificity: Very high
Antibody Cross Reactivity: Almost no significant cross-reactivities detected in HepG2 and HeLa cells, except 27 kDa protein in HeLa; in HepG2 cells, insulin increases 27 kDa protein.
Related Product 1: PAK4-pS474 blocking peptide
Related Product 2: PAK4Subtide - PAK4 protein kinase substrate peptide
Scientific Background: PAK4 is a protein-serine/threonine kinase of the STE group and STE20 family and PAKB subfamily. It is highly expressed and widely distributed in most tested human tissues. It stimulates cell survival, inhibits apoptosis, and also plays a role in cell cycle progression. PAK4 has been implicated in the regulation of cytoskeletal organization, and cell morphology, motility and transformation. Binding of GTP-bound Cdc42 to the autoregulatory region at the N-terminus of PAK4 releases monomers from the autoinhibited dimer, enables phosphorylation at S474 and activation of its phosphotransferase activity. It phosphorylates and inactivates the protein phosphatase SSH1, which leads to increased inhibitory phosphorylation of the actin binding/depolymerizing factor cofilin and stabilization of actin filaments. It phosphorylates LIMK1, a kinase that also inhibits the activity of cofilin. Other substrates include integrin beta5/ITGB5, ARHGEF2, RAN, and BAD. It inhibits apoptosis by preventing caspase-8 binding to death domain receptors in a kinase independent manner. It plays a role in cell-cycle progression by controlling levels of the cell-cycle regulatory protein CDKN1A. PAK4 mutations are associated with increased carcinoma cell motility and influenced inhibitor sensitivity. The protein kinase was found to be necessary for oncogenic transformation of breast cancer cells.
Figure 1. Epitope mapping of PAK4-pS474 antibody with similar phosphopeptides on dot blots.
Figure 2. Identification of phosphosites related to PAK4-pS474.
© Kinexus Bioinformatics Corporation 2017