Collagen Substrate, FITC Conjugate

FITC-labeled bovine type I Collagen Substrate is an excellent substrate for examining collagenase activity. This labelled substrate is highly purified and telo-peptide free. To minimize background levels in collagenase assays, FITC-labeled collagen has been enzymatically pre-treated and further purified by ion-exchange chromatography. 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 flourescence intensity at 520 nm (Emission)/490 nm (Excitation).

Bovine skin_x000D_ _x000D_ We certify that:_x000D_ a) The collagen fluorescein is of animal origin only._x000D_ b) The collagen fluorescein was derived only from animals which were slaughtered in the United States_x000D_ in USDAinspected abattoirs where the animals were subjected to ante- and post mortem inspection_x000D_ and the animals were fit for use in laboratory studies._x000D_ c) The United States if free from foot and mouth disease and rinderpest.

Used for the assay of mammalian collagenase activity. To heat denature collagen for preparing a substrate for gelatinase, transfer collagen solution to a brown glass bottle and heat to 80ºC._x000D_ _x000D_ Collagen-fluorescein may be used as a substrate for the determination of collagenase activity by quantitative fluorescent assay of lysed collagen. Excitation is 490 nm, emission is 520 nm._x000D_ 1) Dialyze at 5°C vs. 0.1 M Tris pH 7.5 containing 0.2 M NaCl._x000D_ 2.) Follow procedure of Baiciet. al. Anal. Biochem., vol 108, pg. 230 (1980)._x000D_ The collagen-FITC is solubilized with collagenase at pH 8.0 at 37°C and absorbance at 494 nm determined in a 10 mm cell. Using € = 7200 at pH 8 at 494 nm;_x000D_ umoles FITC per mg = 0.24.

This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving Humans or animals.

1. Terato, K., et al. 'A rapid assay method of collagenase activity using 14C-labeled soluble collagen as substrate.' Biochim. Biophys. Acta 1976, 445, 753-762_x000D_ 2. Sellers, A., et al. 'Evidence that latent collagenases are enzyme-inhibitor complexes.' Biochem. J. 1977, 163, 303-307_x000D_ 3. Shinkai, H., et al. 'A complex of collagenase with low molecular weight inhibitors in the culture medium of embryonic chick skin explants.' J. Biochem. (Tokyo) 1977, 81, 261-263_x000D_ 4. Shinkai, H. , Nagai, Y., 'A latent collagenase from embryonic Human skin explants.' J. Biochem. (Tokyo) 1977, 81, 1261-1263_x000D_ 5. Murawaki, Y., et al. 'Serum collagenase activity in patients with chronic liver disease.' J. Hepatol. 1993, 328-334_x000D_ _x000D_ 6. Baici, A., Cohen, G., Fehr, K. and Boni, A., Anal. Biochem., 108, 230, (1980).