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smearing of proteins during transfer - (Apr/10/2008 )

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QUOTE (Flour Power @ Apr 22 2008, 03:53 AM)
Out of interest, why are you using PVDF membranes for tris-tricine and nitrocellulose for tris-glycine?

if you know nitrocellulose always works why not just stick to that? It's what i always use with tricine gels.

Our lab historically has used nitrocellulose with tris-glycine gels. With this antibody that I'm working with, we can detect recombinant protein, but not the endogenous protein using our standard methods. The recombinant protein is much more highly expressed and slightly bigger than the native protein. My boss is very adamant that we detect the native protein (it would help our assays a lot if we could detect the real thing), so he asked me to try everything possible to see it which is difficult because its small, membrane associated, not very highly expressed, and we believe that it degrades easily. So, i decided to try tricine gels and PVDF membranes which are supposed to be better for smaller proteins. I had one successful experiment so far, but I honestly don't know if its attributable to using the tricine and PVDF.


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