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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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problem with crosslinker
Posted by: acasanova (IP Hidden, New member, 1)
Date: August 17, 2005 08:08AM
hi, I need some help, may be someone can help me....
the problem is the following, I’m trying to Co-Ip two proteins and I use a crosslinker due that the interaction (I believe) is very weak. To do that I co-transfect both plasmids in HEK293 cells (over night), then apply the crosslinker for half hour and then quench it. The issue is that when I lysate the cells with standard lysis buffer (either triton 1% or RIPA) I get only 10% (aprox) of total protein in comparation when i don't use crosslinker (obviously, using that same amount of cells and plasmids when I carry the co trasfection). Any advice will be welcome!!!! Thanks a lot
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