Rapid Collagenase Assay Kit

The rapid assay method consists of four steps:

1. Activation of latent collagenase in test samples,
2. Reaction with substrate for 10-60 minutes,
3. Denaturation and extraction of degraded products, and
4. Measurement of fluorescence intensity (FI) in the extract.

The enzyme reaction is 10 times faster than substrate gel analysis and much more sensitive than assays using radiolabeled collagen.

Kit includes labeled collagen to perform 100 assays.

FITC-Labeled Collagen

Arthrogen-CIA^® FITC-labeled bovine type I is an excellent substrate for examining collagenase activity.  This labeled substrate is highly purified and telo-peptide free and is supplied lyophilized or in solution.  To minimize background levels in collagenase assays, Arthrogen-CIA^® FITC-labeled collagen has been enzymatically pre-treated and further purified by ion-exchange chromatography.

Arthrogen-CIA^® FITC-labeled collagen can also be used as a substrate for cell culture. Collagen degradation products in the culture supernatants can be directly determined by measuring the fluorescence at 520 nm (Emission)/490 nm (Excitation).

Recombinant Human PMN Pro-Collagenase

Arthrogen-CIA^® r-Human PMN Pro-Collagenase (MMP-8) (see reference 10) is supplied in solution (10 units/vial, 100 units/ml).  This latent pro-collagenase can be used for various purposes such as a reference standard for assaying collagenase activity.  The most critical process in assaying for collagenase is the activation of inactive collagenase, since this varies depending upon the method, reagents, and type of enzyme.  Thus, it is important to use the same reference standard for individual tests. Furthermore, it can be used as a control for testing collagenase inhibitors.

Recombinant Human MMP-13

MMP-13 (collagenase 3) is a newly discovered matrix metalloproteinase found in various tissues such as malignant tumors, osteoarthritic cartilage, rheumatoid synovium and wounds.  MMP-13 production in chondrocytes and synoviocytes is upregulated by stimulation with inflammatory mediators such IL-1, TNF, and retinoic acid.
MMP-13 has been shown to degrade types I and II collagen although the degradation of type II occurs approximately 10 times faster than that of type I collagen.  Although the typical 3/4 and 1/4 fragments were produced as with MMP-1 (collagenase 1), MMP-13 generates a second cleavage of type II collagen that removes three amino acids from the amino terminus of the 1/4 fragment.  MMP-13 also degrades aggrecan; the major proteoglycan of cartilage.

This Arthrogen-CIA^® MMP-13 Kit contains a truncated, human recombinant MMP-13 (rMMP-13) enzyme which cleaves a fluorogenic peptide substrate (included in the kit). This kit is ideal for assessing inhibitors of this novel enzyme. This kit will perform 100 reactions and contains all necessary buffers and reagents.

Pricing

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Last modified: January 03, 2007