Hi Sandip-
The degradation may well be due to proteases in the Dicty lysates, are you using a protease inhibitor cocktail?
best,
Richard Gomer

________________________________________
From: DICTY [[log in to unmask]] on behalf of Sandip Sutariya [[log in to unmask]]
Sent: Thursday, September 15, 2016 12:18 PM
To: [log in to unmask]
Subject: [DICTY] adprt1A (Dd PARP1) protein

Dear Dicty Community,

We are hoping someone might have worked/working on adprt1A gene. We are facing a problem with cloning of full length adprt1A gene. However, we have cloned BRCT domain of it in pGEX4T1 vector and expressed it in E.coli cells. BRCT protein is stable but while performing pull down assay with Dicty lysate, it degrades within few minutes.

Please kindly give your valuable suggestions.

Thanks and best wishes,

Sandip

--
Sandip Sutariya
Senior Research Fellow,
Prof. Rasheedunnisa Begum's Lab,
Dept. of Biochemistry,
Faculty of Science,
The Maharaja Sayajirao University of Baroda.
Vadodara-390002.
Gujarat,India.

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