PARP can be cleaved by caspases and Granzymes. Caspase-3 cleavage generates an 89 kDa band. Granzymes generate 50kDa and 42 kDa bands. All (Full length, 89, 50, 42 kDa Bands) are recognized by PARP-1 (Santa cruz Sc-7150) antibody. Use 15% gel. Good Luck.

Thank you Goodwin G. Jinesh... I am planing to purchase that antibody...

It's also possible that your compound induces automodification of PARP-1 by long and branched ADP-ribose polymers (up to 200 residues in chain) on several sites (up to 25) of the automodification domain. This process gives raise to higher molecular weight PARP-1 forms that do not enter the polyacrylamide gel matrix...

Thank you Daniela Montariello,

You may rite, because after treatment the band was slightly higher than normal band.

Please find the attached image for your reference.

I have some question to you?

1. Like you said automodification of PARP will induce cell death or survival mechanism?

2. How can I detect this kind of PARP?

3. My compound start disappearing the PARP band after 6 h treatment until 72 hrs, but it didn't induce cell death.

4. there are several PARP inhibitor under clinical trial such as Iniparib, olaparib and rucaparib.

5. What is the difference between PARP inhibitor and automodification? Actually, this is very basic question still I want some clarification.

Thanking you.

Sorry its confidential...

Thanks for your suggestion.

I am thinking that there are some other enzyme which can cleave the PARP followed by

Calpain-1~70-40 kD N-terminal fragments

Cathespsin-B and -D 89, 74, 62 kD N-terminal fragments and 55, 44 kD C-terminal fragments

Cathespsin-G 74, 62 N-terminal fragments and 55, 44 kD C-terminal fragments

Granzyme-A 55 kD N-terminal fragment and ~55 kD C-terminal fragment

Granzyme-B 64, 61 kD N-terminal fragments and  54, 42 kD C-terminal fragments

MMP-2 48 kD and 66 kD fragment

This fragments cant be detect by cell signalling antibody.

I would like to know which antybody can detect this kind of fragments ?

and whether this cleavage cause cell death or not?

Hi Mr Pasupathi,

I had run the Western blot to detect the cleavage of PARP in the treated cells using various compounds. After detection, I realized that there were more than 2 cleavage of PARP bands appear in my blot in the treated cells. While the control untreated cells only has one band formed around 80-94kDa range. I am wonder if the band appear in the control untreated cells is not the cleaved PARP band ? Thank you very much for your help.

Hii

I am also getting a lot of cleaved parp bands in my sample without any treatment. The aim is to get only one parp band without any cleavage. How can I achieve this?