CST-Cell signaling technology
Cleaved PARP (Asp214) Antibody (Mouse Specific)
#9544
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Poly(ADP-ribose) polymerase (PARP) is a nuclear DNA binding protein which can detect DNA strand breaks and is involved in base excision repair. Once PARP is cleaved by caspase during apoptosis, its DNA repair function is impaired. Therefore, cleaved PARP has been used as an indicator of apoptosis in Western blot analysis. Using anti-cleaved PARP antibody, Western blot was perfomed to detect induction of apoptosis in HEK293 cells transiently expressing gene X which has been known as an inducer of apoptosis when overexpressed. This showed cleaved PARP detected in cells transiently expressing gene X. However, in cells transfected with a variant form of gene X apoptosis is not induced, thus PARP cleavage was not detected.
Western blot
HEK293 cell for overexpression of gene X and variant form of gene X
Overnight incubation at 4 degrees C, 3% BSA in TBST
5% skim milk in TBST
2 hour incubation at room temperature, 3% BSA in TBST
NA
ECL solution for HRP detction
When HEK293 and empty vector were used as a negative control, no cleaved PARP was observed. Cleaved PARP, which is a indicator of apoptosis, was detected in HEK293 cells transiently expressing gene X which has been known as a inducer of apoptosis when everexpressed. However, HEK293 cells transfected with a variant form of gene X would not induce apoptosis according to level of cleaved-PARP. Actin was used as a loading control.
None
Very specific and sensitive
It only detects the cleaved form of PARP which is an indicator of apoptosis. It is very specific and very sensitive.