I read in several papers, that HUVECs (Shedding of the Matrix Metalloproteinases MMP-2, MMP-9, and MT1-MMP as Membrane Vesicle-Associated Components by Endothelial Cells; doi: 10.1016/S0002-9440(10)64887-0 ) If i would want to measure this shedding in the supernatant after diverse treatments (e.g. LPS) in ELISA, do I have to break the vesicles to release the enzymes or how can i measure the qunatitive change?
Hi Nadia Gallenstein . The easiest way to identify and measure MMPs is gelatin zymography. It can also be semi quantitative.
Hi Nadia,
I agree with Steingrimur about using zymography (at least for MMP2 and 9). Any MMPs encapsulated in exosomes are not likely to be detected by ELISA, unless you treat your supernatant with lipases or use other methods to break the vesicles. You may also want to consider zymography using substrates other than gelatin (i.e. casein etc.) in order to detect MMPs other than gelatinases.