Hi,
I am working on the same project of measuring sequestering on Neutrophiles using MPO Assay. I am a newbie at this method. I have tenatively developed a protocol using o-dianisidine as a subtrate. I have a question: I have a lysis buffer consisting of potassium phosphate, Hexa trimethylammonium Bromide (HTAB), and EDTA. The buffer seems to form a white precipitate after mixing the reagents. I am guessing that the source of the white precipitate is EDTA. Is this normal? Do I resuspend the precipitate before I add to tissue samples?
I also have substrate buffer consisting of potassium phosphate, hydrogen peroxide, and 0-dianisidine diHCl. This buffer also forms a white precipitate. Is this normal as well? I think that the source of the precipitation is the o-dianisidine.
pH for these buffers are 6.0.
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