Chitinase Assay from Cultured Bone Marrow Derived Macrophages
Danielle Worth1, J Philip Nance1, Emma H Wilson1
1Division of Biomedical Sciences, University of California, Riverside, USA.
Bio-Protocol
|July 26, 2016
Summary
This study details an assay to measure chitinase activity in macrophage cell lysates. The assay uses a fluorescently labeled chitin substrate to quantify enzyme function.
Area of Science:
- Biochemistry
- Enzymology
- Cell Biology
Background:
- Chitinases are enzymes that degrade chitin, a key component of fungal cell walls and arthropod exoskeletons.
- These enzymes are crucial in host defense against chitin-containing pathogens.
- Chitinases also play a significant role in the inflammatory processes associated with asthma.
Purpose of the Study:
- To develop and describe a fluorometric assay for detecting chitinase activity.
- To quantify chitinase enzyme function within macrophage cell lysates.
Main Methods:
- Utilizes a 4-methylumbelliferone (4-MU)-labeled chitin substrate.
- Measures the release of 4-MU via hydrolysis of the chitin substrate.
- Employs fluorometric detection to determine enzyme activity levels.
Main Results:
- The assay successfully detects and quantifies chitinase activity.
- Provides a method for measuring enzyme levels in macrophage cell samples.
Conclusions:
- This fluorometric assay is effective for assessing chitinase activity in macrophage lysates.
- The assay facilitates research into the role of chitinases in immunity and inflammatory diseases like asthma.
