Detection of Human Phospho-Chk2 (T68) by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 1 µM camptothecin (CPT) for 1 hour. PVDF membrane was probed with 0.5 µg/mL Rabbit Anti-Human Phospho‑Chk2 (T68) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1626) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-Chk2 (T68) was detected at approximately 64 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda -phosphatase ( lambda -PPase) for 60 minutes. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-Chk2 (T68) in HepG2 Human Cell Line. Phopsho-Chk2 (T68) was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line treated with 1 µM camptothecin using Rabbit Anti-Human Phospho-Chk2 (T68) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1626) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei in treated cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
ATM phosphorylates the Checkpoint kinase on T68 in response to genotoxic insults. Phosphorylation of T68 promotes the formation of Chk2 oligomers. Oligomerization is a critical event in the activation of the Chk2 kinase as it is required for phosphorylation of additional Chk2 residues that lead to full activation and signaling to downstream effectors.
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