Product Name: Brk-pS446+pY447
Product Number: AB-PK547
Size: 25 µg      Price:89.00
      $US
Target Full Name: Protein-tyrosine kinase 6

Target Alias: Brk6; PTK6; Tyrosine-protein kinase 6; FLJ42088; CCDS13524.1; Q13882; ENSG00000101213

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK547

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: S446+Y447

Protein UniProt: Q13882

Protein SigNET: Q13882

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human Brk sequence peptide Cat. No.: PE-04AHP00
Antibody Immunogen Sequence: SFT(pS)(pY)ENP(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues S443 to P450; In the 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.

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: Weak immunoreactivity of a target-sized protein by Western blotting in A549 cells. 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 50-55 kDa.

Antibody Specificity: Very high

Antibody Cross Reactivity: No significant cross-reactivities detected in A549 except EGF increase in 75 kDa protein. Five higher molecular mass cross-reactive proteins (~75 to 250 kDa) in sea star oocytes.

Related Product 1: Brk-pS446+pY447 blocking peptide

Related Product 2: Brk-pY342 phosphosite-specific antibody (Cat. No.: AB-PK548)

Scientific Background: Brk (PTK6) is a protein-tyrosine kinase of the TK group and Src family. It is a non-receptor kinase that is implicated as a regulator of several intracellular signalling pathways involved in cellular differentiation, epithelial maintenance, and tumourigenesis. The activity of Brk appears to be dependent upon the cell type and intracellular localization of the protein. In non-tumour cells, Brk activity promotes cellular differentiation and apoptosis, while in cancer tissue Brk activity sensitizes cells to growth promoting signals leading to increased cell proliferation, anchorage-independent cell survival, and cell migration/invasion. The nuclear form of the protein is thought to be involved in regulating epithelial growth. The 451-amino acid sequence of the Brk protein resembles that of a Src family kinase, containing a SH3, SH2, and a catalytic domain. However unlike Src kinases, the N-terminal domain of the Brk protein does not contain a myristolyation domain, indicating different regualtory mechanisms. Brk mRNA is expressed at high levels in the colon, small intestine, and prostate as well as at low levels in some fetal tissues.
Figure 1. Epitope mapping of BRK-pS446+pY447 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to BRK-pS446+pY447.