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GO
Gail Otulakowski
Hospital for Sick Children
Research fields
Molecular biology
Estimation of Wound Tissue Neutrophil and Macrophage Accumulation by Measuring Myeloperoxidase (MPO) and N-Acetyl-β-D-glucosaminidase (NAG) Activities
Authors:
Puebla Cassini-Vieira
,
Camila Francisco Moreira
,
Mariana Felipetto da Silva
and
Lucíola da Silva Barcelos
,
date:
11/20/2015,
view:
13278,
Q&A:
0
The inflammatory response is essential to the reestablishment of cutaneous homeostasis following injury. In this context, leukocytes arrive at the wound site and orchestrate essential events in the wound healing process. Therefore, the quantification of specific subsets of inflammatory cells in the wound tissue is of considerable interest. The current protocol focus on a quantitative index of neutrophils and macrophages accumulation within skin lesions by measuring the specific activity of the marker enzymes Myeloperoxidase (MPO) and N-acetyl-β-D-glucosaminidase (NAG), respectively. MPO is present in high levels in the azurophilic granules of neutrophils and NAG in lysosomes of activated macrophages. These methods allow the indirect estimation of the abundance of neutrophils and macrophages accumulated into the skin.
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MPO on small tissue biopsies
[Feedback 1] I used this protocol - disruption with glass beads and assay on 96-well plate - on small lung tissue biopsies. It worked very well, much more convenient for large numbers of samples than our previous more traditional method.
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