Elastase, 10 mg

Porcine pancreatic elastase is a serine protease with a molecular mass of 25.9 kDa and an optimal catalytic pH of 8.5. The enzyme hydrolyzes peptide bonds on the carboxyl side of small, hydrophobic amino acids such as glycine, alanine, valine, leucine, and isoleucine, and additionally exhibits activity toward amides and esters, e.g., N-benzoyl-L-alanyl methyl ester.

A defining feature of elastase is its ability to cleave native elastin, a substrate not degraded by trypsin, chymotrypsin, or pepsin. It also hydrolyzes other proteins, including fibrin, casein, hemoglobin, albumin, and denatured collagen. Proteolytic activity can be inhibited by soybean trypsin inhibitor or kallikrein inhibitor, while elastolytic function remains unaffected. Due to this substrate specificity, elastase is frequently used in combination with other proteases for enzymatic cell dissociation, enabling efficient isolation of cells from animal tissues or culture surfaces.

Twice crystallized, supplied as a dialyzed, lyophilized powder.
The enzyme should be filtered through a 0.22 µm filter after reconstitution and prior to use.
Suitable for the isolation of Type II lung cells.
At neutral pH and concentrations >0.25 %, solubility is limited; it is recommended to prepare primary solutions in KCl or alkaline buffers before diluting the enzyme into reaction mixtures or media to compensate for changes in ionic strength and pH. Stable pH range: 4.0–10.4.

One unit of enzyme activity is defined as the amount of enzyme that hydrolyzes 1 mg of Elastin-Congo Red within 20 minutes under the specified conditions.
Elastase activity is assayed using a method adapted from Feinstein et al., Biochem. Biophys. Res. Comm., 50, 1020 (1973), employing the more soluble substrate described by Bieth et al., Biochem. Med., 11, 350 (1974).