Mouse specific cleaved PARP (cell death marker)

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LMP
University of Toronto
Graduate Student

Company:

Cell Signaling

Product Name:

Cleaved PARP (Asp214) Antibody (Mouse Specific)

Catalog Number:

9544

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Programmed cell death (PCD) is a critical process that underlies normal tissues development and serves as a homeostatic mechanism to maintain cell populations. Importantly, dysregulation of PCD has been demonstrated to promote various disease pathologies and is a contributing factor in the development of most cancers. The main form of PCD is known as apoptosis, which results in the activation of numerous proteolytic enzymes termed caspases. Substrates of caspases include a wide range of host proteins that induce cell death upon proteolytic cleavage. This review focuses on the cleavage of PARP-1 that was utilized as a read-out for apoptotic cell death.

Experimental Design and Results Summary

Applications

Western blot

Sample

Mouse embryonic fibroblasts (MEFs)

Primary Incubation

5% skim milk in TBST with anti-cleaved PARP 1/1000- 1 hour at room temperature

Blocking Agent

5% skim milk in TBST- 10 minutes at room temperature

Secondary Incubation

1/10000, room temperature for 1 hour in TBST 5% skim milk

Tertiary Incubation

NA

Detection

HRP- chemiluminescence

Results Summary

This antibody only recognizes the cleaved form of PARP (~89kDa). Apoptosis was achieved only when MEFS were stimulated with apoptotic stimuli (CHX, cycloheximide and TNF) as cleaved PARP was detected.

Additional Notes

NA

Related Categories

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Summary

The Good

Highly specific antibody with no background detection on Western blot.

The Bad

Mouse specific....but works great!

The Bottom Line

Another great antibody from Cell Signaling!!

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