I'm currently developing a serological dipstick test and I have difficulties in generating robustness in my results. I'm using antibody-HRP as my detection with 4CN as a precipitate but I would like to create a SOP to increase the robustness for my assay. But when i replicate my tests I get quite high variation in the intensities in my spots. Would changing into DAB generate more reproducible results? Is it simply impossible to get a good robustness when using an enzyme reaction to visualise primary antibodies?
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