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A modified method allows for correlation between NADPH-diaphorase histochemistry and immunohistochemistry for the
Z Grozdanovic1, G Nakos, B Mayer
1Department of Anatomy, Free University of Berlin, Germany.
Abstract:
Results obtained with the conventional nitro blue tetrazolium salt method for the visualization of the NADPH-diaphorase (NADPH-d) activity of nitric oxide synthase (NOS) are not specific for this particular enzyme, since this activity represents only a fraction of the total cellular NADPH-d pool. Therefore, the standard NADPH-d procedure was modified by performing the incubation in the presence of formaldehyde. Parallel application of the modified NADPH-d staining technique and the indirect immunofluorescence using an antibody against the neuronal isoenzyme (nNOS) on rat, mouse and guinea-pig tissues showed a correlation between histochemical and immunocytochemical staining. It can thus be concluded that the modified NADPH-d procedure allows for a more specific detection of the histochemical nNOS activity than the conventional method.