The Phospho-CHEK2 (Ser516) Antibody is a polyclonal antibody designed to detect endogenous levels of Checkpoint Kinase 2 (CHEK2) protein only when phosphorylated at serine residue 516 (Ser516). This phosphorylation event occurs in response to DNA damage and replication stress, playing a critical role in CHEK2’s activation and downstream signaling .
Key characteristics:
Immunogen: Synthetic peptide derived from human CHEK2 (amino acids 486–535) containing phosphorylated Ser516 .
Applications: Western blot (WB), immunohistochemistry (IHC), immunofluorescence (IF), and ELISA .
CHEK2 is a serine/threonine kinase central to DNA damage checkpoint control. Phosphorylation at Ser516 regulates its activity and interactions:
The antibody has been used to map CHEK2 activation in UVB-irradiated cells, showing caffeine-dependent suppression of ATR/Chk1 pathways .
Identifies CHEK2’s role in stabilizing RAD51 chromatin association during homologous recombination repair .
Mutations in CHEK2 (e.g., Li-Fraumeni syndrome) correlate with breast cancer, sarcomas, and prostate cancer. The antibody aids in assessing CHEK2 phosphorylation status in tumor samples .
Phospho-Ser516 CHEK2 levels are inversely correlated with MDM2-mediated p53 degradation, influencing chemotherapy responses .
Specificity: ≤1% cross-reactivity with non-phosphorylated CHEK2 peptides .
Buffer Composition: PBS with 50% glycerol and 0.02% sodium azide (common across vendors) .
Batch Consistency: Validated using siRNA-mediated CHEK2 knockdown and phosphopeptide competition assays .
Phospho-CHEK2 (Ser516) detection is critical for: