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Title
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Measuring myeloperoxidase activity as a marker of inflammation in gut tissue samples of mice and rat
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Author
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Abstract
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| Myeloperoxidase (MPO) is an enzyme contained in lysosomal azurophilic granules of neutrophils. MPO activity has been shown to correlate with the number of neutrophils in histological sections of the gastrointestinal tract and is therefore accepted as a biomarker of neutrophil invasion in the gut. This protocol describes an easy, cost-effective kinetic colorimetric assay to quantify myeloperoxidase activity in intestinal tissue samples. It is explained using tissue collected in mice but can also be used for other laboratory animals. In a first step, tissue specimens are homogenized using a phosphate buffer containing 0.5% hexadecyltrimethylammonium bromide (HTAB), which extracts MPO from neutrophils. The obtained supernatant is added to a reagent solution containing o-dianisidine dihydrochloride, which is a peroxidase substrate. Finally, the change in absorption is measured via spectrophotometry and converted to a standardized unit of enzyme activity. The assay is illustrated and compared to a commercially available enzyme-linked immunoassay (ELISA), demonstrating that MPO activity does not necessarily correlate with MPO protein expression in tissue samples. |
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Language
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English
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Source (journal)
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Bio-protocol
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Publication
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2023
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DOI
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10.21769/BIOPROTOC.4758
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Volume/pages
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13
:13
(2023)
, p. 1-9
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Article Reference
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e4758
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ISI
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001089325800013
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Pubmed ID
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37456337
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Medium
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E-only publicatie
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Full text (Publisher's DOI)
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Full text (open access)
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