Zymography and reverse zymography for detecting MMPs and TIMPs.
Zymography is the electrophoretic separation of proteins through a polyacrylamide gel containing a proteolytic substrate. After denaturing (but nonreducing) electrophoresis, proteins are renatured and incubated in an appropriate buffer for proteolytic activity. Clear zones of lysis in the stained gel indicated active proteinases. Reverse zymography is a similar technique to detect proteinase inhibitors. After renaturing of proteins, the gel is incubated with metalloproteinases which digest the substrate incorporated into the gel. Inhibitors are shown as dark zones of inhibition against a clear background upon staining.
Department of Biopharmaceutical Sciences, University of California, San Francisco, CA, USA.
Susan P
SP
Hawkes
Hongxia
H
Li
Gary T
GT
Taniguchi
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D020782:Matrix Metalloproteinases / Q000032:analysis*; D013194:Staining and Labeling; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis*
D019714:Tissue Inhibitor of Metalloproteinases; D020782:Matrix Metalloproteinases