Collagenase NB 5 Sterile Grade, CAS No.9001-12-1

CAS: 9001-12-1 Cat. No.: C279009 Número EC: 232-582-9 PubChem CID: 44384985

GRADE & PURITY EnzymoPure™, Sterile, ≥0.10 U/mg

Accession #

P43153,Q46085,Q9X721

Bioactivity

≥0.10 U/mg

Descripción general

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.

Specifications

Product Name

Collagenase NB 5 Sterile Grade, CAS No.9001-12-1

Sinónimos

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

Grado

Sterile, EnzymoPure™

Especificaciones y pureza

EnzymoPure™, sterile, ≥0.10 U/mg

Mecanismos bioquímicos y fisiológicos

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

Bioactividad

≥0.10 U/mg

Número de adhesión

P43153, Q46085, Q9X721

Peso molecular previsto

70–120 kDa

CAS

9001-12-1

Número de la Comisión de Enzimas

3.4.24.3

Tipo de molécula

Enzyme

Almacenamiento y envío

Concentración

≥0.10 U/mg

Reconstitución

It is recommended to prepare a stock solution by dissolving collagenase NB in an appropriate volume of buffer (e.g., HBSS, PBS, Tris, or Ringer’s solution). The prepared enzyme should be sterile-filtered (if not sterile), aliquoted and stored at -20°C. Our stability studies indicate that the enzyme remains stable for up to one year under these conditions, provided that repeated freeze-thaw cycles are avoided.

Condiciones de almacenamiento de almacenamiento

Store at 2-8°C

Enviado en

Wet ice

Estabilidad y almacenamiento

Store at 2-8℃ long term (>1 year). Upon reconstitution, it is recommended to aliquot.

Definición de unidad

The hydrolysis of 1 µmol 4-phenylazobenzyloxycarbonyl-L-prolyl-L-leucylglycyl-L-prolyl-D-arginine per minute at 25℃, pH 7.1.

Objetivos asociados (humanos)

HT-29 (80576 Activities)

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MCF7 (126967 Activities)

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SK-MEL-5 (47095 Activities)

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A549 (127892 Activities)

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Mecanismos de acción

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