胶原酶 NB 5 来源于溶组织梭菌

Collagenase I | Collagenase Type II | (5S,6S,9R,10S,13S,17S,23S,24S,27R,28S,31S,35S)-5,6,9,13,17,23,24,27,31,35-Decamethyl-10,28-dioctyl-2,20-diazanonacyclo[19.15.0.03,19.05,17.06,14.09,13.023,35.024,32.027,31]hexatriaconta-1(21),2,19-triene | microbial c

C279009-1g

1g

EnzymoPure™

≥0.10 U/mg

Collagenases from Clostridium histolyticum are proteolytic enzymes that cleave peptide bonds in the triple helical collagen molecules of human or animal tissue in situ. For this reason collagenases are widely used for isolation of various cell types by tissue dissociation. Collagenase NB 5 Sterile Grade is sterile according to European Pharmacopoeia. Collagenase NB 5 Sterile Grade is a crude collagenase that contains collagenolytic and additional enzymatic activities including clostripain and neutral protease. The balanced ratio of these activities ensures gentle and efficient tissue dissociation. Specification Collagenase activity: ≥0.10 U/mg (PZ acc. to Wünsch) Sterility: must comply Application Collagenase NB 5 Sterile Grade is suitable for dissociation of a broad variety of tissue types. Storage conditions Collagenase NB 5 Sterile Grade is available as a lyophilized powder. It should be stored at +2 to +8℃ in a dry environment. Under these conditions the product is stable until the minimum shelf life stated on the certificate of analysis if repeated opening and closing of the vial is avoided. For storage of solutions please refer to “Stock solution”. Instructions for use: General Collagenase NB 5 Sterile Grade is suitable for isolation of a broad variety of cells from human or animal tissues. Tissue types include adipose tissue, cartilage, skin, placenta, and umbilical cord tissue. It can also be applied in cell culture for passaging, e.g. of embryonic stem cells. Tissue dissociation Recommended starting concentrations for selected applications: Adipose tissue (human, rodent): 0.2 – 0.3 PZ U/ml Cartilage (human, rodent): 0.3 – 0.4 PZ U/ml In general, the appropriate collagenase concentration depends on tissue type and origin as well as on the isolation procedure. Collagenase activity is at an optimum at 37℃ and pH 7.4. Stock solution Collagenase NB 5 Sterile Grade dissolves at a concentration of up to 150 mg/ml in all buffers which are commonly used for cell isolation. The enzyme solution must be constantly stored on ice. Since collagenase and some of the secondary proteases depend on calcium, it is recommended to use a buffer with ≥ 2 mM Ca²⁺. Absolutely no calcium chelating agents (e.g. EDTA) should be present at all. Reconstituted Collagenase NB 5 Sterile Grade can be aliquoted and stored at -20 °C. Aliquots are stable for 1 year if repeated freezing and thawing is avoided. Collagenase NB 5 Sterile Grade is sterile according to Ph. Eur. Therefore, 0.22 µm filtration is not necessary if sterile equipment and buffers are used. If 0.22 µm filtration is required, filters with low protein-binding properties (e.g. cellulose acetate, PVDF, or PES) are recommended. Working solution To prepare a working solution, the stock solution is diluted with buffer to achieve the required collagenase concentration. The working solution must be constantly stored on ice until use. Inactivation and inhibitors The dissociation process can be reduced, e.g. by cooling down or dilution of the enzyme solution. Collagenase is reversibly inactivated at high pH values and irreversibly inactivated at low pH values. Inhibitors of collagenase include cysteine or chelating agents like EDTA. Important note Collagenase NB 5 Sterile Grade is intended for research use only.

Clostridial collagenases are among the most efficient degraders of eukaryotic collagen known; saprophytes use collagen as a carbon source while pathogens additionally digest collagen to aid in host colonization. Has both tripeptidylcarboxypeptidase on Gly-X-Y and endopeptidase activities; the endopeptidase cuts within the triple helix region of collagen while tripeptidylcarboxypeptidase successively digests the exposed ends, thus clostridial collagenases can digest large sections of collagen. Catalytic activity: Digestion of native collagen in the triple helical region at Xaa-|-Gly bonds. With synthetic peptides, a preference is shown for Gly at P3 and P1', Pro and Ala at P2 and P2', and hydroxyproline, Ala or Arg at P3'.

≥0.10 U/mg

天然

70–120 kDa

溶组织梭菌(Clostridium histolyticum)