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C6885

Sigma-Aldrich

Collagenase from Clostridium histolyticum

suitable for release of physiologically active rat epididymal adipocytes, Type II, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid

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Synonym(s):
Clostridiopeptidase A
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

biological source

Clostridium histolyticum

Quality Level

form

powder

specific activity

≥125 CDU/mg solid
0.5-5.0 FALGPA units/mg solid

suitability

suitable for release of physiologically active rat epididymal adipocytes

application(s)

diagnostic assay manufacturing

shipped in

wet ice

storage temp.

−20°C

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General description

Clostridium histolyticum is a pathogenic clostridium that produces collagenase. This is a mixture of enzymes containing collagenase, non-specific proteases and clostripain.

Application

Collagenase may be used:
  • for the preparation of arterial tissue for the study of advanced glycosylation end products (AGE)
  • for use along with other proteases for the disaggregation of human tumor, mouse kidney, human brain, lung epithelium and many other tissues.
  • in liver and kidney perfusion studies, digestion of pancreas, and isolation of nonparenchymal hepatocytes
  • for the preparation of viable hepatocytes from rat liver and for the isolation of fat cells from rat adipose tissue

Biochem/physiol Actions

Collagenase is activated by four gram atom calcium, per mole enzyme. It is inhibited by ethylene glycol-bis(β-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, β-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline.Collagenase enzymes and neutral protease plays an important role in the effective release of cells from tissue. Collagenase recognizes the sequence -R-Pro-8-X-Gly-Pro-R-, where X is most often a neutral amino acid.
Collagenase is activated by four gram atom calcium per mole enzyme. It is inhibited by ethylene glycol-bis(beta-aminoethyl ether) - N, N, N′,N′-tetraacetic acid, beta-mercaptoethanol, glutathione, thioglycolic acid and 8-hydroxyquinoline.

Caution

As supplied, this product is stable for one year at -20°C. There is no loss in FALGPA or protease activity in 30 days at 37°C, 50°C and -20°C. Solutions of crude collagenase are stable if frozen quickly in aliquots (at 10 mg/mL) and kept frozen at -20°C. Further freeze-thaw cycles will damage the solution. The product retains 100% activity over 7 hours when held on ice.

Unit Definition

One collagen digestion unit (CDU) liberates peptides from collagen from bovine achilles tendon equivalent in ninhydrin color to 1.0 μmole of leucine in 5 hours at pH 7.4 at 37 °C in the presence of calcium ions. One FALGPA hydrolysis unit hydrolyzes 1.0 μmole of furylacryloyl-Leu-Gly-Pro-Ala per min at 25°C. One Neutral Protease unit hydrolyzes casein to produce color equivalent to 1.0 μmole of tyrosine per 5 hr at pH 7.5 at 37°C. One Clostripain Unit hydrolyzes 1.0 μmole of BAEE per min at pH 7.6 at 25°C in the presence of DTT.

Preparation Note

Solutions are typically prepared at 1-2 mg/mL in TESCA buffer (containing 50 mM TES, 0.36 mM Calcium chloride, pH 7.4 at 37°C.
This product also contains clostripain, nonspecific neutral protease, and tryptic activities.

pictograms

Health hazardExclamation mark

signalword

Danger

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

target_organs

Respiratory system

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

dust mask type N95 (US), Eyeshields, Faceshields, Gloves


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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D. Schomberg and M. Salzmann
Enzymes, 1 (1991)
Allah Nawaz et al.
Nature communications, 8(1), 286-286 (2017-08-19)
Adipose tissue resident macrophages have important roles in the maintenance of tissue homeostasis and regulate insulin sensitivity for example by secreting pro-inflammatory or anti-inflammatory cytokines. Here, we show that M2-like macrophages in adipose tissue regulate systemic glucose homeostasis by inhibiting
C M Posavad et al.
Mucosal immunology, 10(5), 1259-1269 (2017-01-05)
Local mucosal cellular immunity is critical in providing protection from HSV-2. To characterize and quantify HSV-2-reactive mucosal T cells, lymphocytes were isolated from endocervical cytobrush and biopsy specimens from 17 HSV-2-infected women and examined ex vivo for the expression of
O Matsushita et al.
Journal of bacteriology, 181(3), 923-933 (1999-01-28)
Clostridium histolyticum collagenase contains a number of different active components. Previously we have shown that colH encodes a 116-kDa collagenase (ColH) and a 98-kDa gelatinase. We purified a different 116-kDa collagenase (ColG) from the culture supernatant and sequenced its gene
Seglen, P.O.
Methods in Cell Biology, 13 (1976)

Articles

Organoid culture products to generate tissue and stem cell derived 3D brain, intestinal, gut, lung and cancer tumor organoid models.

Protocols

To measure collagenase activity, N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala is used in a continuous spectrophotometric rate determination at 345 nm. Collagenase hydrolyzes collagen peptide bonds.

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