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Protocol for cell surface protein co-IP

E. coli surface antigen co-IP

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#1 milawa

milawa

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Posted 17 April 2012 - 09:59 PM

Hi there,
I have a bacterial cell surface protein which we think is part of a larger protein complex and I have been asked to prove this by adding antibody, directed to my protein of interest, to whole bacterial cells, wash away any unbound antibody and then solubilise the membranes and perform a co-IP. I don't see anything bound to the beads expect antibody using my current methods (however a normal co-IP shows the complex intact)
I can see the protein on the cell surface by immunofluorsecence experiments and protease shaving experiments.

Any suggestions or protocols would be appreciated.

Thanks !

Edited by milawa, 17 April 2012 - 10:14 PM.






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