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Author Topic:   pull down assay
seeker
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posted 02-12-2001 03:02 PM           Edit/Delete Message Reply w/Quote
I am having some problems with my pull down assay.I have successfully cloned my bait
protein into pCI-NEO vector and prey protein into pCDNA3 vector. I have also sequenced the junctions to ensure they are in frame. I did a co-transfection into cos-1 cells and did a lysis reaction after 12hrs, 24 hrs and 36hrs. When I do a western, I got exactly the same bands for the lanes containing my samples and lanes with the empty vectors alone. I am using anti-myc and anti-flag antibodies. Please give me your precious advice.
Thank you

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