Wed Jun 16 05:28:17 EST 1993
You may try 25 mM Borate, pH 10 instead of TRIS, but values much about pH 10 could hydrolyse PAA. Setting up an acidic system and running the proteins cathodically could also be a way out. I have heard about but never tried myself. E.g. Glycine, pH 5, without a discontinuos buffer system. Be sure that your protein is soluble at this pH.
If you intend to detect your protein by antibody reaction, be careful with Triton ! It may destroy epitopes. Test this by a dot blot assay before running a Triton-gel. Triton might also disaggregate non-covalently attached protein complexes in several cases.
Good luck and let me know how you solved this problem !
Peter Hammerl, c/o hartla at dsbgpx.edvz.sbg.ac.at
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