We would like to measure the MPO activity from myocardial tissue from control Vs treated groups. We have the tissue lysate in RIPA buffer. Is it possible to measure the MPO activity from these lysate which was stored in -80 deg C. Many protocol says the tissue should be homogenized in MPO assay buffer and fresh. Is colorometric activity assay is the best assay or any alternative assays (e.g. Immunoblot of MMP 2/9 etc.) available. We do not have the intact tissue so can not perform sectioning and staining now.
Hi Narendra.
Heart lysates in RIPA buffer should normally work for MPO assay. However, it depends on the kit. We have tried ELISA with tissue lysate in RIPA buffer and it worked always.
Alternative strategies depends on your aim. If you want to measure MPO activity, then MPO assay kits or ELISA are the best option. If you are interested in MPO mass, then immunoblot of MMP2/9 or MPO should be sufficient.
For activity measurements, I would suggest to go for ELISA than colorimetric assays.
All the best,
Chintan
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