Detection of peroxidase by the print technique in thin-layer isoelectric focusing.

摘要: Abstract About twenty secondary substrates were screened for the detection of peroxidase by print technique following thin-layer isoelectric focusing. The best results obtained with o-toluidine, guaiacol, o-phenylenediamine, and o-dianisidine. Detection horseradish was optimal when paper buffered at pH 5, impregnated a methanolic solution containing 1% urea-peroxide 1–2% substrate. Uniform staining enzyme zones (compared to metachromatic many other substrates) high color stability on storage obtained. o-Toluidine guaiacol less sensitive factor 10–100 than o-phenylenediamine o-dianisidine, which belong most chromogens. With two latter as little 0.001–0.01 μg could be detected technique. gave almost perfect white backgrounds contrasting zones, whereas o-dianisidine yielded moderately stained, but still acceptable, background. Peroxidase described has been found suitable also separation methods, e.g., gel filtration.