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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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To solve problem
Posted by: Chethana P. Gowda (IP Hidden, Unregistered user, )
Date: April 7, 2005 07:12AM
Hi,
I am trying to immunoprecipitate 26kd protein from the mouse epidermis so for i am not successful. Please could you suggest to solve this problem.And i am following the procedure from the Santa cruz protocol. Thanks Chethana P. Gowda
Re: To solve problem
Posted by: bassamfahmawi (IP Hidden, Regular member, 42)
Date: April 12, 2005 08:30AM
Hi,
Do you detect your protein in a Western blotting experiments using this antibody? If not, this might mean that your protein is membane (Check for transmembane domain(s) on [www.psort.org]) or cytosceleton associated you need to use detergents (SDS denaturing) in your lysis buffer to solublize your protein. However, if you do so you need to remove the SDS later on from your lysis buffer before you apply for IP, otherwise the SDS will impose its denaturing effect on your antibody as well used in the IP. Please refer to the IP procedure I have posted earlier on this forum (IMAC). Good Luck!
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