A spectrophotometric collagenase assay |
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| Authors: | A Nethery J G Lyons R L O'Grady |
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| Institution: | 1. Institute of Biochemistry of Biologically Active Compounds, National Academy of Sciences, Grodno, Belarus;2. School of Medical Sciences, Bialystok, Poland;3. Lviv National Ivan Franko University, Lviv, Ukraine;4. Institute of Cell Biology, National Academy of Sciences, Lviv, Ukraine;1. Food Technology Department, UTPV-XaRTA, Agrotecnio Research Center, University of Lleida, Alcalde Rovira Roure 191, 25198 Lleida, Spain;2. School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore;3. Animal Facilities, Campus Ciències de la Salut, Facultat de Medicina, University of Lleida, C/Montserrat Roig 2, 25008 Lleida, Spain;1. University of Iowa, Iowa City, IA;2. PinMed, Inc., Pittsburgh, PA;3. University of Pittsburgh, Pittsburgh, PA;4. Yale University, New Haven, CT |
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| Abstract: | A quantitative collagenase assay using Coomassie blue staining and microtiter spectrophotometry is described. Collagen is gelled and dried onto the bottom of microwells as substrate, washed, incubated with samples, washed again, and then stained. Absorbance at 590 nm increases linearly with increasing amounts of collagen in the range 5-40 micrograms. Bacterial and mammalian collagenases can be detected within 2 h, and 10 ng of bacterial collagenase may be detected in 16 h. For simple screening applications, activity may be detected by eye. The assay is safe, simple, fast, economical, and sensitive. |
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