Collagenases assays

Mammalian collagenases and the matrix metalloproteinases Collagenase assays Collagen-based assays Peptide-based assays... 

Collagenase assays were carried out in 50 mM Tricine, 0.2 M NaCl, 10 mM CaClj (pH 7.5) at 30°C using rat type I collagen as substrate. 

Stack and Gray have described a convenient, continuously recording, fluorescent assay for rabbit collagenase and gelatinase based on the hydroly-... 

A number of studies have used synthetic collagenase inhibitors in physiological assays with live tissue or whole animals for the purposes of either establishing the role of a collagenase in a physiological event, or as a test of the efficacy of the inhibitor. Since these represent two of the most important... 

Samples were taken from each solution after incubation for calcium determination. Samples of the collagenase incubations were assayed for hydroxyproline. 

Fig. 8. Variation of the skin wound half-life with degradation rate R (in collagenase) of collagen-GAG matrix. The half-life is the time required for a wound to contract to 50% of the original area. The degradation is in empirical units, which are defined in terms of an in vitro assay. A somewhat arbitrary broken vertical line is drawn near R = 140 enzyme units. This line shows the level of degradation rate above which the half-life of matrices rapidly drops to the level of the ungrafted wound. The horizontal scale is logarithmic [79]...

Garbisa, S., Tryggvason, K., Foidart, J. M. and Liotta, L. A. (1980b) Assay for radiolabeled type IV collagen in the presence of other proteins using a specific collagenase. Anal. Biochem. 107,187-192. 

A EXPERIMENTAL FIGURE 7-19 Oocyte expression assay is useful in comparing the function of normal and mutant forms of a channel protein. A follicular frog oocyte is first treated with collagenase to remove the surrounding follicle cells, leaving a denuded oocyte, which is microinjected with mRNA encoding the channel protein under study. [Adapted from I R Smith, 1988, Trends Neurosci. 250.]... 

Collagenase production and release are partly responsible for the joint destruction that characterises human rheumatoid arthritis. Triterpenes from the lupane and a-amyrin groups have been studied in vitro to examine their effects on the release of the arthritic joint degradative enzyme collagenase using the rat osteosarcoma. This test and the rat synovial granuloma of adjuvant arthritis are similar both models are based on connective tissue tumours with bone-invasive properties. The pentacyclic triterpenes assayed have been shown to possess general antiproteolytic effects that can explain the anti-arthritic effects in adjuvant arthritis in rats [71,103]. 

Zll. Zucker, S., Mancuso, P., DiMassimo, B., Lysik, R. M., Conner, C., and Wu, C.-L., Comparison of techniques for measurement of gelatinase/type IV collagenases Enzyme-linked immunoassays versus substrate degradation assays. Clin. Exper. Metastasis 12, 13—23 (1994). 

Yoshioka, H., et al. (1987). An Assay of Collagenase Activity Using Enzyme-linked Immunosorbent Assay for Mammalian Collagenase, Anal. Biochem. 166 172-177. 

The crude fish enzyme extracts were prepared as per the method of Baranowski et al. (1984), and stored in ice for use in the pressure treatments and subsequent enzyme assays. The spectrophotometric methods of Hummel (1959) and Erlanger et al (1961) were used to assay for chymotrypsin-like and trypsin-like enzyme activities using BTEE and BAPNA as substrates, respectively. Cathepsin C activity was assayed using gly-phe-NA as substrate (Lee et al, 1971), and collagenase activity in the fish extracts were assayed as per the method of Wunsch and Heidrich (1963). Protein content of the crude enzyme extracts from fish was determined by the method of Hartree (1972). 

The degradation rate was decreased by crosslinking CG matrices in 0.25% glutaraldehyde. Degradation rates in collagenase were assayed in... 

An early version of the in vitro collagenase degradation assay was based on measurement of stress relaxation of a strip of test polymer immersed in a standardized collagenase bath. Previously, Tobolsky had used such a device to study the chemical stress relaxation of natural rubber in the presence of ozone. In spite of its immediate utility as a screening probe in our early research this experimental arrangement was eventually replaced by a more reproducible assay. The latter is an adaptation of a colorimetric procedure originally developed by Mandl and coworkers. ... 

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