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B20224

Sigma-Aldrich

Collagenase - Elastase blend

Tissue dissociation blend, lyophilized powder, suitable for cell culture

Synonym(s):

Clostridiopeptidase A, Clostridium histolyticum, Elastase, Pancreatopeptidase E, collagenase, collagenase blend, enzyme blend, tissue dissociation blend

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About This Item

UNSPSC Code:
12352204
NACRES:
NA.71

Quality Level

assay

(enzymatic)

form

lyophilized powder

concentration

1 mg/mL (Collagenase)

technique(s)

cell culture | mammalian: suitable

shipped in

dry ice

storage temp.

-10 to -25°C

Related Categories

General description

The Collagenase-Elastase 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 Elastase: a serine protease that also hydrolyzes amides and esters. It is produced in the pancreas as an inactive zymogen and activated in the duodenum by trypsin. While elastase will hydrolyze a wide variety of protein substrates, it is unique among proteases in its ability to hydrolyze native elastin, a substrate not attacked by trypsin, chymotrypsin or pepsin. Elastin, a matrix component of tissues involved in stretching processes, is found in highest concentrations in the elastic fibers of connective tissues. Elastase is frequently used to dissociate tissues which contain extensive intercellular fiber networks. For this purpose, it is usually used with other enzymes such as collagenase, trypsin, and chymotrypsin. Elastase is the enzyme of choice for the isolation of Type II cells from the lung.

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. Elastase: hydrolyses elastin, the specific protein of elastic fibers, and digests hemoglobin, casein and fibrin. It cleaves protein at the carboxyl side of small hydrophobic amino acids such as Ile, Gly, Ala, Ser, Val, and Leu. The enzyme also hydrolyzes amides and esters such as N-Benzoyl-L-alanine methyl ester. The pH optimum is found to be 8.0-8.5. It does not require any activator, but it is inhibited by diisopropyl fluorophosphate, phenylmethanesulfonyl fluoride, α2-macroglobulin, α1-antitrypsin, sulfonyl fluorides and p-dinitrophenyl diethylphosphate and high salt concentrations. It cleaves protein at the carboxyl side of small hydrophobic amino acids such as Ile, Gly, Ala, Ser, Val, and Leu. The enzyme also hydrolyzes amides and esters such as N-Benzoyl-L-alanine methyl ester.

Preparation Note

For a working concentration of: Collagenase of 1 mg/ml, Elastase 3 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

signalword

Danger

Hazard Classifications

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

target_organs

Respiratory system

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3


Certificates of Analysis (COA)

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Articles

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

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