C3-22-BC | Collagenase Type III, CLS III

C3-22-BC
  
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      Overview

      Replacement Information
      Description
      OverviewCells of animal tissue are attached to each other by a complex matrix of proteins, glycoproteins, lipides, glycolipides, and mucopolysaccharides. To isolate individual cells or to start primary cultures, the delicate matrix must be partially broken down without destroying the cell or its surface. The treatment of tissue with Collagenase affects a careful, selective reduction of the intercellular matrix and does not influence the growth ability of the cells. The raw Collagenase offered by us is a mixture of different proteolytically efficient enzymes. For optimal results, an exactly adjusted mixture of the proteolytic enzymes is necessary. For this purpose we offer 4 different types of Collagenase, CLS I, CLS II, CLS III, and CLS IV. The last one is normally applied with other enzymes like trypsin, elastase, or hyaluronidase. The Trypsin or Trypsin/ EDTA generally used in cell cultures attacks the matrix only slowly, causing moreover irreversible damages of the released cells.

      Specifications:

      Enzyme blends of collagenase, clostripain, with tryptic and proteolytic activities from Clostridium histolyticum. This preparation is recommended for liver, lung, adipose, and adrenal tissue. Specific activity is 125 to 250 Mandl units per ml of powder substance.

      Recommendations:

      Actual enzyme concentrations should be 0.1 to 0.2% (w/v), depending on the delicate nature of the tissue substrate. This is valid for an activity of about 160 Mandl units per milligram of dry substance. On other specific activities the concentration of use has to be modified correspondingly. The indication of the specific activity is related to the respective method of determination. The determination of activity according to Mandl is difficult to designate due to the high molecular collagen substrate. As it is a biological product, collagenase is subject to natural variations from charge to charge. Thus, we recommend to previously test deliverable charges concerning their suitability for the desired purpose and to optimize the process conditions (concentration, incubation time, and temperature).

      Definitions:

      • Collagenase activity according to Mandl: 1 unit releases 1 µmol amino acids determined with ninhydrin and calculated as leucin from native collagen within 5 hours (+37°C and pH 7.5).

      • Collegenase activity according to Wünsch and Heidrich:
      1 PZ (4-phenyl-azobenzyl-oxycarbonyl) unit catalyses the hydrolysis of 1 µmol of water soluble chromo-peptide PZ-Pro-Leu-Gly-Pro-D-Arg per minute (+25°C/pH 7.1). Resulting PZ-L-Pro-Leu can be determined.
      Catalogue NumberC3-22-BC
      Brand Family Calbiochem®
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      Ship Code Blue Ice Only
      Storage +2°C to +8°C
      Do not freeze Ok to freeze
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