Source: Bacillus polymyxa
Neutral protease (Dispase®) is a non-mammalian animal origin free (AOF) metallo, neutral protease, purified by methods developed at Worthington. Its mild proteolytic action makes the enzyme especially suitable for the preparation of primary cells and secondary (subcultivation) cell culture, since it is gentle on cell membranes. This protease is also used as a secondary enzyme in cell isolation and tissue dissociation applications, commonly used with collagense.
Characteristics of Neutral Protease (Dispase®) from Bacillus polymyxa:
Molecular Weight: 36 kda.
pH Optimum: Stable over a wide pH range: 4.0-9.0, optimum pH 5.9-7.0.
Stability/Storage: Stable at 2-8°C for 12 months. Aliquot and store at -20°C after reconstitution with water or commonly used balanced salt solutions or media.
Unit Definition: One Unit releases one micromole of Folin positive amino acids, measured as tyrosine, at 37°C, pH 7.5, using casein as the substrate.
Specificity: Non-specific cleavage of peptide bonds containing leucine and phenylalanine.
Activators: Divalent cations including Ca2+, Mg2+, Mn2+, and Fe2+
Inhibitors: EDTA (1mM), EGTA, 1-10-phenanthroline and heavy metals
Application(s): Commonly used to separate skin epidermis from dermis leaving intact epithelial sheets and stem cell, hepatocyte and other cell isolation applications. However, due to the diversity of the variables involved, exact isolation conditions should be determined empirically for each cell/tissue application.