Product Name:
Chk2-pT68
Product Number:
ab-pk581
Target Full Name: Checkpoint protein-serine kinase 2
Target Alias: Cds1; CHEK2; Chk2 checkpoint homologue (S. pombe); HuCds1; LFS2; PP1425; RAD53; RP11-436C9.1; bA444G7; CCDS13843.1; ENSG00000183765
Product Type Specific: Chk2 (CHEK2) protein kinase phosphosite-specific antibody
Antibody Code: PK581
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T68
Protein UniProt: O96017
Protein SigNET: Chk2
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human Chk2 (CHEK2) sequence peptide Cat. No.: PE-04AJY99
Antibody Immunogen Sequence: TVS(pT)QEL(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues T65 to L71; in the N-terminal portion of the kinase before the FHA domain. T68 phosphorylation stimulates phosphotransferase activity, and induces interactions with Chk2, MDC1, NBS1 and Plk1. This is one of the minor in vivo phosphorylation sites in Chk2 (≥3 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). Chk2 is known to be phosphorylated at this site in vitro by ATM, ATR, Chk2 (CHEK2), DNAPK (PRKDC), Plk1 (PLK), TTK, and ZAK (MLTK).
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: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus Macaque | Rat | Mouse.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 60-68 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in A431 cells and sea star oocytes.
Related Product 1: Chk2 (65-71) pT68 antibody blocking peptide (Cat. No.: PE-04AJY99)
Related Product 2: Chk2Selectide kinase substrate peptide (Cat. No.: PE-01BGT95)
Related Product 3: Chk1 S280 phosphorylation antibody (Cat. No.: AB-PK577)
Related Product 4: Chk1 S317 phosphorylation antibody (Cat. No.: AB-PK578)
Related Product 5: Chk1 S345 phosphorylation antibody (Cat. No.: AB-PK579)
Related Product 6: Chk2 T383 phosphorylation antibody (Cat. No.: AB-PK580)
Related Product 7: Chk2 T68 phosphorylation antibody (Cat. No.: AB-PK119)
Scientific Background: Chk2 (CHEK2) is a 65 kDa protein-serine/threonine kinase of the CAMK group and RAD53 family. It is required for cell cycle arrest, activation of DNA repair, and the promotion of apoptosis in response to double-stranded breaks in DNA. It features an N-terminal SQ/TQ cluster domain (SCD), a Forkhead-associated (FHA) domain for protein binding, and a C-terminal kinase domain. The forkhead-associated protein interaction domain essential for activation in response to DNA damage. It is rapidly phosphorylated at Thr-68 by MLTK in response to replication blocks and DNA damage, and activated through autophosphorylation. The response to DNA damage occurs in an ataxia telangiectasia mutated (ATM)-dependent manner. It is phosphorylated at Thr-68 by ATM and ATR. It is also activated by phosphorylation at Ser-19, Thr-26, Ser-28, Ser-33, Ser-35, Thr-383, Thr-387 and Ser-516. The effects of Chk2 are mediated through the phosphorylation and inhibition of various downstream effectors, including CDC25A, CDC25B, and CDC25C. Chk2 phosphorylates CDC25C at Ser-216 and inhibits this phosphatase (blocking entry into mitosis), p53 at Thr-18 and Ser-20 (inducing cell cycle arrest in G1), and BRCA1 (maintaining survival after DNA damage). Twelve splice variant isoforms have been described for human Chk2. High expression is observed in testis, spleen, tonsils, colon and peripheral blood leukocytes, and is notably absent in brain and spinal cord. Orthologues of Chk2 are amongst the most highly conserved protein kinases in animals, plants, fungi and unicellular eukaryotes. Expression of wild-type Chk2 leads to increased p53 stabilization after DNA damage, whereas expression of a dominant-negative Chk2 mutant abrogated both phosphorylation of p53 on S20 and p53 stabilization. Loss of function mutants of ChK2 can cause Li-Fraumeni syndrome, a highly penetrant familial cancer phenotype often found with p53 mutations as well. Familial mutations also associated with prostate and breast cancers, and mutations also occur in a variety of sporadic cancers and cell lines. This description may include information annotated by UniProt and/or Google AI.