Merck, B20222-1VL, Collagenase - Hyaluronidase blend

General description
The Collagenase-Hyaluronidase blend is a tissue dissociation enzyme cocktail for cell isolation. The tissue dissociation process consists in the release of functional and viable cells from animal tissue, with minimal impact on the cell’s viability. The main reagent used for the tissue dissociation process is Collagenase. Collagenases are enzymes that break down the native collagen that holds animal tissues together. The second enzyme found in our blend is Hyaluronidase. Hyaluronidase is a hyaluronic acid-metabolizing enzyme, who degrades Hyaluronic Acid. Hyaluronic Acid increase tisue and membranes permeability and is one of the main components of the extracellular matrix, contribuing significantly to cell proliferation and migration. Combination of Collagenase and Hyaluronidase is usually used for the isolation of cells from the extracellular matrix.
Biochem/physiol Actions
Collagenase: Effective release of cells from tissue requires the action of collagenase enzymes. Collagenase is activated by four-gram atom calcium (Ca2+) per mole enzyme. The pH optimum is 6.3-8.8. The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Collagenase treatment can cause some cells to die. Typically, concentrations varying from 0.1 to 5 mg/mL are used for digestion. The duration of reaction varies from 15 minutes to several hours and yields satisfactory cell dissociation without causing too much cell death. Hyaluronidase: degrades hyaluronan and randomly cleave β-N-acetylhexosamine-(1−4)-glycosidic bonds in hyaluronic acid, chondroitin, and chondroitin sulfates.
Preparation Note
For a working concentration of Collagenase of 1 mg/ml and an enzymatic activity of Hyaluronidase 100 units/ml, reconstitute the lyophilized powder with 10 ml of Hank’s Balanced Salt solution with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate (#H6648)