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C0130

Sigma-Aldrich

Collagenase from Clostridium histolyticum

Collagenase from Clostridium histolyticum
1 of 1 reviewers received a sample product or took part in a promotion

for general use, Type I, ≥125 CDU/mg solid

Synonym(s):

Clostridiopeptidase A

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About This Item

CAS Number:
9001-12-1
Enzyme Commission number:
3.4.24.3 (BRENDA, IUBMB)
EC Number:
232-582-9
MDL number:
MFCD00130830
UNSPSC Code:
12352204
NACRES:
NA.54
Pricing and availability is not currently available.

Quality Level

200

form

lyophilized powder

specific activity

≥125 CDU/mg solid

mol wt

68,000 to 125,000

optimum pH

6.3-8.8

solubility

TESCA buffer (50 mM TES, 0.36 mM Calcium chloride, pH 7.4): soluble 1-2 mg/mL at 37 °C

storage temp.

2-8°C

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General description

Collagenase is a protease which cleaves the triple-helical protein called collagen. There are three types of tissue collagenases, and these belong to the matrix metalloproteinases (MMP) family.[1] Collagenase obtained from Clostridium histolyticum has a very strong activity, as it digests collagen from both ends, at temperatures as low as 4-10°C.[2] Crude collagenase mixtures contain two major enzyme types namely, collagenase and clostripain.[3]

Application

Collagenase from Clostridium histolyticum has extensive use in biological studies, where it is used to prepare isolated cell suspensions.[3] This product is suitable for the disaggregation of human tumor, mouse kidney, human adult and fetal brain, lung and many other epithelia tissues. It has also been shown to be effective in liver and kidney perfusion studies, digestion of pancreas, isolation of nonparenchymal rat liver cells and hepatocyte preparation. Collagenase has also been used in the preparation of arterial tissue for the study of Advanced Glycosylation End Products. This enzyme has been tested for the release of heptatocytes at a concentration of approximately 1 mg/mL. Concentrations for digestion range from 0.1 to 5 mg/mL.

Biochem/physiol Actions

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.
The collagenase product is a mixture of enzymes secreted by C. histolyticum, with different products differentiated by the relative ratios of the 10-18 components found in the secreted enzymes. The main components are two collagenases, clostripain, and a neutral protease. The synergistic action of these enzymes degrade collagen and other intracellular materialThe action of both collagenase enzymes and the neutral protease is necessary for effective release of cells from tissue. Various types of collagen are the natural substrates for collagenase.

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

This product is an equivalent to first 40% ammonium sulfate fraction of Mandl, I., et al., J. Clin. Invest., 32, 1323 (1953). 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.

also commonly purchased with this product

Product No.
Description
Pricing

inhibitor

Product No.
Description
Pricing

Pictograms

Health hazardExclamation mark
GHS08,GHS07

Signal Word

Danger

Hazard Statements

H315,H319,H334,H335

Hazard Classifications

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

Target Organs

Respiratory system

Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Certificates of Analysis (COA)

Lot/Batch Number
0000412370
0000412370
0000398572
0000367708
0000363709

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Konstantin Khalturin et al.
Nature ecology & evolution, 3(5), 811-822 (2019-04-17)
Cnidarians are astonishingly diverse in body form and lifestyle, including the presence of a jellyfish stage in medusozoans and its absence in anthozoans. Here, we sequence the genomes of Aurelia aurita (a scyphozoan) and Morbakka virulenta (a cubozoan) to understand
Amanda P Smith et al.
Frontiers in microbiology, 9, 1554-1554 (2018-07-26)
Mathematical models that describe infection kinetics help elucidate the time scales, effectiveness, and mechanisms underlying viral growth and infection resolution. For influenza A virus (IAV) infections, the standard viral kinetic model has been used to investigate the effect of different
Amber M Smith et al.
Scientific reports, 6, 38703-38703 (2016-12-16)
Secondary bacterial infections increase morbidity and mortality of influenza A virus (IAV) infections. Bacteria are able to invade due to virus-induced depletion of alveolar macrophages (AMs), but this is not the only contributing factor. By analyzing a kinetic model, we
Aline Wuidart et al.
Nature cell biology, 20(6), 666-676 (2018-05-23)
The mammary gland is composed of basal cells and luminal cells. It is generally believed that the mammary gland arises from embryonic multipotent progenitors, but it remains unclear when lineage restriction occurs and what mechanisms are responsible for the switch
Meagan Ita et al.
Journal of biomechanical engineering (2019-06-19)
Abnormal loading of a joint's ligamentous capsule activates nociceptive afferent fibers, which reside in the capsule's collagenous matrix alongside fibroblast-like synoviocytes (FLS) and transmit pain to the DRG. This study integrated FLS into a DRG-collagen gel model to mimic the
View All Related Papers

Protocols

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

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.

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

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.

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

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.

Enzymatic Assay of Collagenase (EC 3.4.24.3) using FALGPA (N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala)

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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Questions

1–8 of 8 Questions  

  1. How can I check the activity of product C0103?

    1 answer

    1. The minimum activity of this product is 125 units per milligram. The specific method used to determine the activity of this material is considered proprietary. However, a geneeral activity protocol is available at the following link:
      https://www.sigmaaldrich.com/US/en/technical-documents/protocol/protein-biology/enzyme-activity-assays/enzymatic-assay-of-collagenase-using-n-3-2furylacryloyl-leu-gly-pro-ala?srsltid=AfmBOoq8fzW3O4ksOh8nr0Xe0DToP1T4FK6HnZuy5XtgfWRllrpdEHKV

      Please see the following link to review a sample or lot specific Certificate of Analysis:
      https://www.sigmaaldrich.com/product/sigma/c0130#product-documentation

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  2. Can collagenase be added to dissolve natural collagen fibers in dissolvable sutures like catgut? Also, is item C0130 suitable for this purpose?

    1 answer

    1. Upon verification, there is currently no collagenase product tested with catgut suture as a substrate. However, literature suggests that digestion of dissolvable sutures (collagen or catgut) is achievable using collagenase. Additionally, one might consider adding pepsin for the hydrolysis of catgut, as suggested by a publication where pepsin (pH 1.6) was used alongside collagenase (pH 7.4) to replicate an in vivo environment in an in vitro hydrolysis setting.

      Reference: Biomaterials. 1992;13(7):448-54. doi: 10.1016/0142-9612(92)90165-k. Title: Degradation of collagen suture in vitro and in vivo PMID: 1633219

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  3. What is the Department of Transportation shipping information for this product?

    1 answer

    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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  4. Is Product C0130, Collagenase, sterile?

    1 answer

    1. This product does not consist of a sterile preparation of Collagenase. Upon reconstitution, solutions should be sterile filtered through a 0.22 micron filter.

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  5. What type of Collagenases are present in Product C0130, Collagenase,?

    1 answer

    1. This Collagenase is obtained from the culture filtrate of Clostridium histolyticum. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68 - 130 kDa. It does not contain any Matrix Metalloprotease.

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  6. How should Product C0130, Collagenase, be prepared for use in tissue digestions?

    1 answer

    1. For use in tissue culture applications, Collagenase can be solubilized in calcium-free solutions such as Hanks Balanced Salts (H2387) or Earl's' Balanced Salt Solution (E6267).

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  7. What is the activity of the lot number of Product C0130, Collagenase, which I have?

    1 answer

    1. The activity and specific activity for each lot number can be obtained from the certificate of analysis.

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  8. How should I prepare a stock solution of Product C0130, Collagenase?

    1 answer

    1. Stock solutions of crude Collagenase may be prepared at 10 mg/ml of Collagenase in TESCA buffer (50 mM TES, 0.36 mM calcium chloride, pH 7.4 at 37°C) and are stable if frozen quickly in aliquots and kept frozen at -20°C. Freeze-thaw cycles will damage the enzyme.

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Reviews

1 of 1 reviewers received a sample product or took part in a promotion

Active Filters

  1. Glasgow, UK
    • Reviews 2
    • Vote 1
    5 out of 5 stars.

    Clostridium histolyticum

    Good product. Usefull for cell isolation from heart.

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