Anti-PARP antibody [E102] (ab32138)

How much protein from jurkat cell lysate was loaded onto the western blot shown on your datasheet?

ANSWER:

We loaded 15 ug of protein from Jurkat onto Western blot.

I hope this is helpful. Please contact us again if you have any further questions.

We are obtaining a band at 48Kda with this PARP antibody. Bands are usually expected at around 110kDa, and products from ICE family members, such as caspase 3 and 7are usually 25 and 85kDa. Please explain the 48kDa band.

ANSWER:

Thank you for your telephone enquiry yesterday.

I have done a literature search to investigate the different cleavage products of PARP. There is a reference in which analysis of bovine PARP reveals three sites with high homology to known MMP-2 cleavage recognition sequences. Their experiments show MMP-2 degradation products of PARP corresponding to those theoretically predicted. Western blots for PARP after MMP-2 cleavage reveal loss of PARP as well as appearance of a <48 kDa product. These products appear in a concentration-dependent manner along with a corresponding loss of the 116 kDa PARP full length protein band.

FASEB Journal. 2004;18:690-692.) Matrix metalloproteinase-2 (MMP-2) is present in the nucleus of cardiac myocytes and is capable of cleaving poly (ADP-ribose) polymerase (PARP) in vitro. Jennifer Kwan et al.

At website address: http://www.fasebj.org/cgi/content/full/18/6/690?ck=nck

I hope this helps to explain your results. I can suggest doing a further literature to obtain more information on this.

If you are still concerned about the results you have obtained, please do not hesitate to complete the following questionnaire. This will provide me with your protocol information so I can investigate this case further.

http://www.abcam.com/index.html?section=western&pageconfig=technical&mode=questionaire

If you have any further questions, please do not hesitate to contact us again. Good luck with your experiments.