Cell Signaling Technology
Phospho-Chk1 (Ser345) Rabbit mAb Antibody
2348L
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Chk1 is a key effector kinase that controls cell cycle progression after DNA damage. Activation of Chk1 relies on the phosphorylation of Ser 317 and Ser 345 by ATR. Here I tested the UV-induced phosphorylation of Chk1 pS345 and confirmed that phosphorylation is mediated by ATR using an ATR specific inhibitor.
Western Blot
Human HeLa cancer cell line
1:1000 dilution in TBS-T / 5% nonfat milk. 3 hour incubation at room temperature
TBS-T / 5% nonfat milk
1:5000 of anti-rabbit-HRP
NA
Bio-Rad ChemiDoc
This antibody specifically recognized the damage-induced phosphorylation of Chk1. Total Chk1 blot is used as control for the loading of Chk1.
None
Specific
Not superb sensitivity, but still acceptable
Better to hybridize for at least 2 hours at room temperature or overnight at 4 degree to get strong signal.