Detection of Human Phospho-Chk1 (S317) by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or exposed (+) to 50 J/m2 UV-C for the indicated time. PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human/Mouse/Rat Phospho-Chk1 (S317) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2054) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-Chk1 (S317) was detected at approximately 56 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda -phosphatase ( lambda -PPase) for 1 hour. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-Chk1 (S317) by Simple WesternTM.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 50 J/m2 ultraviolet light (UV) for 2 hours, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Chk1 (S317) at approximately 60 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Chk1 (S317) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2054). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
The Chk1 checkpoint kinase is an integral member of a signaling cascade that controls cell cycle progression. In response to genotoxic or replicative stress, Chk1 is phosphorylated by ATM or ATM-related kinases (ATR) at S317. In turn, Chk1 phosphorylates downstream effectors, such as p53 or the Cdc25 phosphatases to halt cell cycle progression and allow time for repair of incurred damage.
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