Collagenase/Elastase

Application Note: Elastase is assayed using a method adapted from that of Feinstein et al., Biochem. Biophys. Res. Comm., 50, 1020 (1973) and using the more soluble substrate of Bieth et al., Biochem. Med., 11, 350 (1974). 1 SucAla3NA unit is approximately equivalent to 6 elastin digestion units. Aqueous liquid suspensions should be aseptically handled to avoid bacterial contamination. Due to the viscous nature of the aqueous suspension the vial should be rinsed to recover contents. One Unit cleaves one micromole of N-succinyl-L-alanyl-L-alanyl-L-alanine-p-nitroanilide per minute at 25 °C,  pH 8.0. The collagenase assay is a modification of Mandl wherein collagenase is incubated for five hours with native collagen and the extent of collagen breakdown is determined using the Moore and Stein, JBC, 176, 367, (1948) colorimetric ninhydrin method. Collagenase is typically used at concentrations from 0.05 % to 0.5 % (w/v) in balanced salt solutions such as Hank's, Earle's and others. Specific conditions for reactivity should be optimized by the end user.

Synonyms: Clostridium histolyticum, Bacterial collagenases, collagenase, Porcine pancreatic elastase, Protease, ColH, ColG, Chymotrypsin-like elastase family member 1, Elastase-1

Background: Collagenase preparations contain several isoforms of two different collagenases, a sulfhydryl protease, clostripain, a trypsin-like enzyme, and an aminopeptidase. This combination of collagenolytic and proteolytic activities is effective at breaking down intercellular matrices, the essential part of tissue dissociation. Crude collagenases are widely used in enzymatic primary cell isolation and tissue dissociation procedures. Most researchers employ either crude collagenase preparations such as Types 1, 2, 3, and 4 or chromatographically purified collagenase, the latter usually combined with secondary enzymes such as elastase, hyaluronidase, etc. Elastase is a serine protease that also hydrolyzes amides and esters. It is produced in the pancreas as an inactive zymogen, and activated in the duodenum by trypsin. The following information applies to porcine elastase. While elastase will hydrolyze a wide variety of protein substrates, it is unique among proteases in its ability to hydrolyze native elastin, a substrate not attacked by trypsin, chymotrypsin or pepsin. Soybean trypsin inhibitor and kallikrein inhibitor suppress proteolytic but not elastolytic activity. Collagenase is ideal for researches focused in Stem Cell and Biomarker Research.

Gene Name: CELA1, colH, colG

Reconstitution Volume: 10.0 mL

Reconstitution Buffer: Restore with deionized water (or equivalent)

Buffer: None

Stabilizer: None