Catalogue

Background
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).

Synonyms: Extracted from bovine skin

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

Product
FITC-labeled bovine type I Collagen provided as a 1 mg/ml solution in 0.01M acetic acid.

Product Form: FITC

Purity: >95%

Concentration: 1 mg/ml

Applications
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 80°C. 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. 1) Dialyze at 5°C vs. 0.1 M Tris pH 7.5 containing 0.2 M NaCl. 2.) Follow procedure of Baici et. al. Anal. Biochem., vol 108, pg. 230 (1980). 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; umoles FITC per mg = 0.24.

Functional Analysis: Activity Assay

Storage
Product should be stored at -20°C. Aliquot to avoid freeze/thaw cycles

Product Stability: Stable in acidic buffer when stored at -20°C, however slowly degrades when stored in under neutral conditions.

Shipping Conditions: Ship at ambient temperature, freeze upon arrival

Caution
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. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Exalpha Biologicals accepts no liability for any inaccuracies or omissions in this information.

References
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 2. Sellers, A., et al. 'Evidence that latent collagenases are enzyme-inhibitor complexes.' Biochem. J. 1977, 163, 303-307 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 4. Shinkai, H. & Nagai, Y., 'A latent collagenase from embryonic human skin explants.' J. Biochem. (Tokyo) 1977, 81, 1261-1263 5. Murawaki, Y., et al. 'Serum collagenase activity in patients with chronic liver disease.' J. Hepatol. 1993, 328-334 6. Baici, A., Cohen, G., Fehr, K. and Boni, A., Anal. Biochem., 108, 230, (1980).