87-kDa PARP activity

Melissa Greeve gliftor at hotmail.com
Thu Oct 4 04:03:29 EST 2001


Hi Araluen

I worked for a long time (well, months) doing westerns for PARP as an
indicator of apoptosis (could only detect the uncleaved protein, even
in apoptotic control cell lysates..). I'd just started then, and had
huge problems with westerns, so I subsequently hate that protein...
um... no, it was a learning experience. I keep finding new
publications about PARP and its function in the cell..

Anyway, I don't know too much about this, so I won't offer too many
suggestions, just to say that proteins in my westerns run lower than
expected, due to differences in the marker and protein sample
buffers.. But I gather your 120kD band is obviously the whole PARP
(and not your 100kD band)? I will (hopefully) be starting kinase
assays soon, so I'd be interested to hear of reasons why proteins
become denatured/inactive/modified during their capture/precipitation.

Melissa Greeve
PhD Student
University of Western Australia




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