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Blotting of native protein gels
Posted by: Scaffoldman (IP Hidden, New member, 2)
Date: February 20, 2006 01:37AM
Hi guys,
I have expressed a putative biotin-binding protein and I am trying to show that it can still bind biotin. What I have in mind is to separate the recombinant protein on a native gel, transfer it to membrane and then detect the protein on the blot by letting it bind biotinylated HRP followed by ECL detection. I have found several protocols for native gel electrophoresis but so far, could not find anything for electrotransfer or blotting of native gels to membrane. Has anyone got a working protocol for this? Also, would it be possible to detect the protein in-gel and if so, how do I prevent the protein from leaching out during incubation and washing steps? Appreciate any help you can offer. Thanks! Scaffoldman
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