AI Article Synopsis
- In situ zymography is a technique for detecting enzymatic activity in tissue, but traditional fixation methods can hinder this process.
- The study demonstrates that using ethanol- and zinc-buffered fixative (ZBF) allows for the preservation of both tissue morphology and enzymatic function, providing better results than frozen sections.
- Although enzymes can be extracted from ethanol- and ZBF-fixed tissues, the yield is generally lower than from unfixed tissues, and extraction efficiency depends on the buffer used.
Article Abstract
In situ zymography is a method for the detection and localization of enzymatic activity in tissue sections. This method is used with frozen sections because routine fixation of tissue in neutral-buffered formalin inhibits enzyme activity. However, frozen sections present with poor tissue morphology, making precise localization of enzymatic activity difficult to determine. Ethanol- and zinc-buffered fixative (ZBF) are known to preserve both morphological and functional properties of the tissue well, but it has not previously been shown that these fixatives preserve enzyme activity. In the present study, we show that in situ zymography can be performed on ethanol- and ZBF-fixed paraffin-embedded tissue. Compared with snap-frozen tissue, ethanol- and ZBF-fixed tissue showed stronger signals and superior morphology, allowing for a much more precise detection of gelatinolytic activity. Gelatinolytic enzymes could also be extracted from both ethanol- and ZBF-fixed tissue. The yield, as analyzed by SDS-PAGE gelatin zymography and Western blotting, was influenced by the composition of the extraction buffer, but was generally lower than that obtained from unfixed tissue.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2796612 | PMC |
| http://dx.doi.org/10.1369/jhc.2009.954354 | DOI Listing |
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