Cell Signaling Technology
PARP (46D11) Rabbit mAb
#9532
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I used PARP (46D11) Rabbit mAb for Western Blot at the suggested dilution (1:1000) in TBST 1X / 5% Not fat dry milk, at overnight incubation time. Best image has been acquired at highest sensitivity, 1m57s450ms of exposure time. I was able to detects endogenous levels of total full-length PARP-1 (≃116 kDa).
Western Blot
MDA-MB-231, MDA-MB-436, HELA, SUM-149, SUM-1315, JURKAT, PSN1, MIA PaCa-2
5% Not fat milk / TBST 1X, for 2 hours
Not fat milk
Primary antibody (PARP Rabbit mAb), dilution 1:1000 in 5% Not fat milk / TBST 1X, overnight in cold room.
Secondary antibody Goat (Anti-rabbit), dilution 1:5000 in 5% Not fat milk / TBST 1X, 1 hour at room temperature.
Chemiluminescence
PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (1). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (6).Cell Signaling Technology - PARP (46D11) Rabbit mAb detects endogenous levels of total full-length PARP-1 (116KDa), but also the large fragment (89 kDa) produced by caspase cleavage at Asp214. Within this experiment, PARP (46D11) Rabbit mAb helped me in find the change in expression of this protein on different cancer cells line tested.
(1) PMID: 1549180 DOI: 10.1038/356356a0 ; (6) PMID: 9837934 DOI: 10.1074/jbc.273.50.33533
Not fat milk is good as blocking agent.
Antibody has a great signal (clean and with high intensity).
Antibody with high selectivity/sensitivity, highly recommended.