Does anyone have a protocol recommendation for digesting a collagen matrix with MMP-1?
My Dear, Alex Mol
Thermal and mechanical manipulation with proteinases could separate the chains
Hello Alex,
Do you want to assess the resistance to enzymatic degradation?
If the collagen matrix has low content of water (sponge, tissue graft, cell layer) weight loss assessment can be a good option. Check this paper: http://pubs.acs.org/doi/abs/10.1021/acsbiomaterials.5b00563
If not, another option is to measure the released collagen in the medium with hydroxyproline assay (very laborious), Sircol collagen assay, BCA assay or other colorimetric fast assay.
Normally we carry out this experiment in Tris Buffer with CaCl2 and at 50 U/ml of MMP-8 or 250 U/ml of MMP-1, 37C, orbital shaking at 120 - 150 rpm. In the near future, my group will publish an article with this information.
Hope it helps,
Hector
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