High salt and antibody binding

tmorris at uhnres.utoronto.ca tmorris at uhnres.utoronto.ca
Tue Oct 8 13:04:59 EST 2002


Are there any experts on the practical aspects of immunocyt? If so I thank you in advance for advice on this one.

Our lab has a hand-me-down recipe for antibody buffer, for staining imobilized isolated neurones for fluorescent microscopy. It's basically a tris based buffer which includes 500mM NaCl. No one can offer a rational explanation for this, nor can I find anything similar on the web or in books. I've used a borate buffer with a more normal osmolarity (300 mOsmol) and the staining was just as good. I suspect PBS would work also, which is what most books reccomend. 
Such high salt might neutralize static charges on IgG, but why would this help in binding epitope. Surely, the closer to physiological conditions the better. Am I missing somthing? The person who designed this buffer must have had a good reason and I don't want to just ditch a protocol without a clear understanding of the whys and wherefores.


Thanks


Rubic



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