Skip to Content
Merck
All Photos(1)

Key Documents

B20223

Sigma-Aldrich

Collagenase - DNase I - Dispase II blend

Tissue dissociation blend, lyophilized powder, suitable for cell culture

Synonym(s):

Clostridiopeptidase A, Clostridium histolyticum, DNase I, Deoxyribonuclease I, Deoxyribonucleate 5′-oligonucleotido-hydrolase, Dispase II, Protease from Bacillus polymyxa, collagenase, collagenase blend, enzyme blend, tissue dissociation blend

Sign Into View Organizational & Contract Pricing


About This Item

UNSPSC Code:
12352204
NACRES:
NA.71

Quality Level

form

lyophilized powder

specific activity

≥2 units/mL Dispase II

concentration

0.1 mg/mL (DNase)
1 mg/mL (collagenase)

technique(s)

cell culture | mammalian: suitable

shipped in

dry ice

storage temp.

-10 to -25°C

General description

The Collagenase-Deoxyribonuclease I-Dispase II blend is a tissue dissociation enzyme cocktail for cell isolation. The tissue dissociation process consists in the release of functional and viable cells from animal tissue, with minimal impact on the cell’s viability. The main reagent used for the tissue dissociation process is Collagenase. Collagenases are enzymes that break down the native collagen that holds animal tissues together. The second enzyme found in our blend is Deoxyribonuclease I (DNase I). DNase I, is an endonuclease of the DNase family, with a specific role for the degradation of DNA. DNase I is one of the most well characterized endonucleases of mammalian origin. During tissue dissociation, parts of the cells are lysed resulting in a release of DNA. Monomolecular DNA may cause clumping of cells, so addition of DNase I to our blend leads to a degradation of this extracellular DNA, thereby avoiding the loss of cells from undesired clumping. The third enzyme found in our blend is Dispase II. Diapse II is a non-mammalian animal origin free. Its mild proteolytic action makes the enzyme especially suitable for the preparation of primary cells and as a secondary enzyme in cell isolation and tissue dissociation applications. It is commonly used to separate skin epidermis from dermis leaving intact epithelial sheets and stem cell, hepatocyte and other cell isolation applications. This collagenase blend allow is a significantly less harmful cell isolation preventing unwanted cell clumping without cell membrane damage.

Biochem/physiol Actions

Collagenase: Effective release of cells from tissue requires the action of collagenase enzymes. Collagenase is activated by four-gram atom calcium (Ca2+) per mole enzyme. The pH optimum is 6.3-8.8. The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Collagenase treatment can cause some cells to die. Typically, concentrations varying from 0.1 to 5 mg/mL are used for digestion. The duration of reaction varies from 15 minutes to several hours and yields satisfactory cell dissociation without causing too much cell death. Hyaluronidase: degrades hyaluronan and randomly cleave β-N-acetylhexosamine-(1−4)-glycosidic bonds in hyaluronic acid, chondroitin, and chondroitin sulfates.

Preparation Note

For a working concentration of: Collagenase of 1 mg/ml, Deoxyribonuclease I of 0.1 mg/ml and Dispase II of ≥ 2 units/ml, reconstitute the lyophilized powder with 10 ml of Hank’s Balanced Salt solution with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate (#H6648).

Pictograms

Health hazardExclamation mark

Signal Word

Danger

Hazard Statements

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 3


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’.

Already Own This Product?

Find documentation for the products that you have recently purchased in the Document Library.

Visit the Document Library

Articles

Discover pre-mixed collagenase enzyme blends with DNase I, Dispase II, Elastase, and Hyaluronidase and gently dissociate animal tissues in vitro.

Discover pre-mixed collagenase enzyme blends with DNase I, Dispase II, Elastase, and Hyaluronidase and gently dissociate animal tissues in vitro.

Discover pre-mixed collagenase enzyme blends with DNase I, Dispase II, Elastase, and Hyaluronidase and gently dissociate animal tissues in vitro.

Discover pre-mixed collagenase enzyme blends with DNase I, Dispase II, Elastase, and Hyaluronidase and gently dissociate animal tissues in vitro.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service