Hello,
I made a cell lysate with a gram-positive bacterium using lysozyme and sonication. I need to inactivate the lysozyme because we are using the lysate in cell-stimulation assays and do not want the lysozyme to influence the cellular response. Inactivation by heat seems to be the way to go, however, we need to maintain the integrity of bacterial lipids. Has anyone had experience with inactivating lysozyme? What temperature and how long was necessary? Will heating at this temp destroy lipids?
Thank you!
Which lysozyme did you use since the answer will vary by which enzyme you used? Hen egg lysozyme for example is pretty stable until about 70 degrees C, which may be too high for your lipids. Have you considered using another approach to making your lysate, for example freeze-thaw cycles and sonication instead of adding lysozyme.
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