Direct comparison of the sensitivity of enzyme histochemical and immunohistochemical methods: cathepsin B expression in human colorectal mucosa.
Immunohistochemical localization of the proteinase cathepsin B has been compared directly with localization of cathepsin B activity with a catalytic (enzyme) histochemical method. The 2 approaches demonstrate principally different aspects of an enzyme. The immunohistochemical method localizes the enzyme protein whether it is active or not whereas the catalytic method visualizes the functionally active enzyme only. Sensitivity of both approaches to localize low amounts of enzyme protein or activity has never been compared. In the present study, we show that cathepsin B protein has a wider distribution pattern than cathepsin B activity in human colorectal mucosa, which means that inactive cathepsin B protein is present. With respect to sensitivity of the methods, it is shown that cathepsin B protein could only be demonstrated properly when strong signal amplification was applied by using Nanogold with silver enhancement, whereas activity could be demonstrated with a simple and direct fluorogenic histochemical assay. It is concluded that catalytic histochemical methods are relatively simple methods for the localization of activity of enzymes in tissues and cells and that their sensitivity is high in comparison with immunohistochemical methods.
Bleeker
F E
FE
Academic Medical Center, University of Amsterdam, Department of Cell Biology and Histology, The Netherlands.
Hazen
L G
LG
Köhler
A
A
Van Noorden
C J
CJ
D002384:Catalysis; D002401:Cathepsin B / Q000096:biosynthesis*; D003106:Colon / Q000378:metabolism* / Q000473:pathology; D004798:Enzymes / Q000378:metabolism; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D009092:Mucous Membrane / Q000378:metabolism* / Q000473:pathology; D012007:Rectum / Q000378:metabolism* / Q000473:pathology; D012680:Sensitivity and Specificity; D013997:Time Factors
D004798:Enzymes; D002401:Cathepsin B