Product Name:
Csk-pY184
Product Number:
ab-pk589
Target Full Name: C-terminus of Src tyrosine kinase
Target Alias: CYL; MGC117393; MPK-2; CCDS10269.1; ENSG00000103653
Product Type Specific: Csk protein kinase phosphosite-specific antibody
Antibody Code: PK589
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y184
Protein UniProt: P41240
Protein SigNET: Csk
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human Csk sequence peptide Cat. No.: PE-04AGJ95
Antibody Immunogen Sequence: DEF(pY)RSG(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues D181 to G187; in the region between the SH2 domain and the protein kinase catalytic domain. Y184 phosphorylation stimulates phosphotransferase activity. This is the major in vivo phosphorylation site in Csk (≥40 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).
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 of a target-sized protein by Western blotting in A431 cells. Medium immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus Macaque | Dog | Rat | Mouse | Chicken | Frog | Zebra fish.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 50-52 kDa.
Antibody Specificity: Medium
Antibody Cross Reactivity: Strong ~76 kDa cross-reactive protein detected in A431, HeLa and Jurkat cells, as well as a strong ~180 kDa cross-reactive protein in Jurkat cell.
Related Product 1: CSK (181-187) pY184 antibody blocking peptide (Cat. No.: PE-04AGJ95)
Related Product 2: CskSubtide kinase substrate peptide (Cat. No.: PE-01BGY95)
Scientific Background: CSK is a 50 kDa non-receptor protein-tyrosine kinase of the TK group and Csk family. It is a predominantly cytoplasmic, and acts as a crucial negative regulator of Src family kinases (SFKs such as Src, Fyn, Lck). It inhibits cell growth, differentiation, and migration by phosphorylating a conserved C-terminal tyrosine residue on SFKs (Tyr-527 of Src), inducing an inactive conformation. It also mediates immune responses by suppresses signalling through T-cell and B-cell receptors. A Y184F mutation leads to inhibited phosphorylation of CSK. Its activity can be decreased by 2/3 with a Y304F mutation. PRKACA induced phosphorylation of CSK can be inhibited with a CSK mutation S364A. It features an SH2 and an SH3 domain, which allow it to be recruited to sites of active SFKs to inhibit them. It is essential for normal cell differentiation and development. Unlike other Src-family kinases, CSK is not membrane-anchored. It must be recruited to the plasma membrane by adapter proteins like Cbp/PAG1 or paxillin to reach its targets. Binding to these adapters also increases its catalytic activity. Mutations or dysregulation of CSK are associated with autoimmune diseases, specifically Systemic Lupus Erythematosus (SLE). Absence of CSK causes severe developmental defects, such as failure in neural tube closure, due to uncontrolled Src activity. This description may include information annotated by UniProt and/or Google AI.