High Sensitivity Detection with PARP Antibody

Cancer System Imaging (Diagnostic Imaging Division)
MD Anderson Cancer Center - University of Texas
Lab Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

PARP (46D11) Rabbit mAb

Catalog Number:

#9532

Clone Number:

N/A

Info:Supplier Page View Product Specs

Browse Similar Products: PARP Antibodies

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).

Experimental Design and Results Summary

Applications

Western Blot

Sample

MDA-MB-231, MDA-MB-436, HELA, SUM-149, SUM-1315, JURKAT, PSN1, MIA PaCa-2

Primary Incubation

5% Not fat milk / TBST 1X, for 2 hours

Blocking Agent

Not fat milk

Secondary Incubation

Primary antibody (PARP Rabbit mAb), dilution 1:1000 in 5% Not fat milk / TBST 1X, overnight in cold room.

Tertiary Incubation

Secondary antibody Goat (Anti-rabbit), dilution 1:5000 in 5% Not fat milk / TBST 1X, 1 hour at room temperature.

Detection

Chemiluminescence

Results Summary

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.

DOI or PMID #

(1) PMID: 1549180 DOI: 10.1038/356356a0 ; (6) PMID: 9837934 DOI: 10.1074/jbc.273.50.33533

Additional Notes

Not fat milk is good as blocking agent.

Image Gallery

Summary

The Good

Antibody has a great signal (clean and with high intensity).

The Bad

N/A

The Bottom Line

Antibody with high selectivity/sensitivity, highly recommended.

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