Cell Signaling Technology
Phospho-Chk1 (Ser345) Rabbit mAb
2348L
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Chk1 is phosphorylated on Ser 317 and Ser 345 by ATR protein kinases after DNA damage. Phosphorylation of Chk1 at S317 and S345 is a hallmark of activation of the cell cycle checkpoint. The UV-induced phospho-Chk1 S345 was examined using immunofluorescent microscopy.
Immunofluorescence
HeLa human cancer cell line
Dilution 1:50 in PBS/0.05% Tween20/3% BSA blocking buffer. Overnight incubation at 4C
3% BSA / 0.05% Tween20 in PBS
Cy3-conjugated anti-Rabbit 2nd antibody at dilution of 1:250 and 1hr incubation at room temperature
N/A
Fluorescent microscope (NIKON)
UV irradiated HeLa cells showed strong and clear Phosphorylation of Chk1 S345 signal, whereas control (undamaged) cells did not.
The samples were fixed in 100% MeOH for 20 minutes at -20 degrees
Very specific
Need to use very high concentration (1:50)
Great antibody for IF study of phospho-Chk1 S345.