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Figure 2 | Molecular Brain

Figure 2

From: Fluorometric immunocapture assay for the specific measurement of matrix metalloproteinase-9 activity in biological samples: application to brain and plasma from rats with ischemic stroke

Figure 2

Rat and human MMP-9 specifically and efficiently binds to mouse monoclonal anti-MMP-9. Recombinant MMP-9 activity and specificity was measured. (A) Dose response of rat and (B) human recombinant MMP-9 was determined (n = 3–5). (C) Activity of free versus immunocaptured rat and (D) human MMP-9. Free MMP-9 was not bound to the MMP-9 antibody but instead was free-floating. Rat and human free vs. immunocaptured MMP-9 activity did not differ (p > 0.05, n = 4). All relative fluorescent units were normalized to a substrate control with 0 ng MMP-9. (E) Rat and (F) human recombinant MMP-2 was incubated with or without antibody to check the specificity of the MMP-9 antibody. No signal was detected with rat MMP-2 and very little signal was detected with human MMP-2 in the immunocapturing condition whereas free active MMP-2 did have fluorescent activity (n = 2–3). (G) Immunoblot for anti-MMP-9 specificity to MMP-9. Rat and human recombinant MMP-9 (92 kDa) and MMP-2 (72 kDa) were probed with the mouse monoclonal anti-MMP-9 then detected with a secondary antibody against mouse (in green). To confirm presence of MMP-2, an anti-MMP-2 antibody was used and counterstained (in red). 10 ng of rat MMP-9 was not enough protein to have significant binding with the MMP-9 antibody.

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Molecular Brain

ISSN: 1756-6606