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Journal of Histochemistry and Cytochemistry, Vol. 49, 1189-1190, September 2001, Copyright © 2001, The Histochemical Society, Inc.

BRIEF REPORT

Stability of Nitroblue Tetrazolium-based Alkaline Phosphatase Substrates

Gary B. Smejkala and Charles A. Kaulb
a Cleveland Clinic Foundation, Department of Clinical Pathology, Cleveland, Ohio
b Lerner Research Institute, Department of Cell Biology, Cleveland, Ohio

Correspondence to: Gary B. Smejkal, Proteome Systems, Inc., 14 Gill Street, Woburn, MA 01801. E-mail: gary.smejkal@proteomesystems.com

This report demonstrates the stability of NBT substrate after multiple exposures to alkaline phosphatase. Perhaps more important than the ability to reuse substrates, the report provides some insight into the mechanisms by which tetrazoliums are reduced and evidence for the formation of an intermediary product, i.e., a half-formazan that is reduced more rapidly. (J Histochem Cytochem 49:1189–1190, 2001)

Key Words: alkaline phosphatase, 5-bromo-4-chloro-3-indolyl, phosphate, formazan, nitroblue tetrazolium chloride, von Willebrand factor, Western blotting


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