Tissue Desintegration Enzymes

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Collagenase (CLS), raw

Collagenase from Clostridium histolyticum
Cells of animal tissue are attached to each other by a complex matrix of proteins, glycoproteins, lipides, glycolipides, and mucopolysaccharides. To isolate individual cells or to start primary cultures, the delicate matrix must be partially broken down without destroying the cell or its surface. Collagenase are the enzymes of choice for this application, if dissolved in balanced salts. Actual enzyme concentrations should be 0.1 to 0.2% (w/v), depending on the delicate nature of the tissue substrate.

Definitions:
Collagenase activity according to Mandl: 1 unit releases 1 µmol amino acids with ninhydrin, determined and calculated as leucin, from native collagen within 5 hours (+37°C and pH 7.5).
Collagenase activity according to Wünsch and Heidrich: 1 PZ-unit catalyzes the hydrolysis of 1 µmol of the water soluble chromopeptide PZ-Pro-Leu-Gly-Pro-D-Arg per minute (+25°C / pH 7.1). Resulting PZI-L-proline (PZ = 4-Phenyl-azobenzyloxycarbonyl) can be determined.

References: Tissue Culture, Methods and Applications (ed. P.F. Kruse and M.K. Patterson) Academic Press, New York (1972)

CLS Type 1:
Enzyme blending from collagenase, clostripain, with tryptic and proteolytic activities, from Clostridium histolyticum . This preparation is recommended for liver, lung, adipose, and adrenal tissue. Specific activity is 125 to 250 Mandl units per milligram of powdered substance.

CLS Type II:
Enzyme blending from collagenase, clostripain, with tryptic and proteolytic activities, from Clostridium histolyticum . This preparation shows a high clostripain activity with the tryptic activity close to type I. Suitable for liver, bone, thyroid, heart, and salivary gland tissue. Specific activity is 125 to 250 Mandl units per milligram of powdered substance.

CLS Type III:
Enzyme blending from collagenase, clostripain, with tryptic and proteolytic activities, from Clostridium histolyticum . This preparation has a very low proteolytic and a normal collagenase activity. Suitable for mammary gland tissue. Specific activity is 125 to 250 Mandl units per milligram of powdered substance.

CLS Type IV:
Enzyme blending from collagenase, clostripain, with tryptic and proteolytic activities. Recommende for the isolation of Langerhans' island cells. Specific activity is 125 to 250 Mandl units per milligram of powdered substance.

Pack/Density

Unit

Cat.no.


Collagenase Storage: 24 months at +2 - +8°C	- / -	100 mg	C 1-28

	- / -	1 g	C 1-22



Collagenase Storage: 24 months at +2 - +8°C	- / -	100 mg	C 2-28

	- / -	1 g	C 2-22



Collagenase Storage: 24 months at +2 - +8°C	- / -	100 mg	C 3-28

	- / -	1 g	C 3-22



Collagenase Storage: 24 months at +2 - +8°C	- / -	100 mg	C 4-28

	- / -	1 g	C 4-22

Papain

Pack/Density

Unit

Cat.no.


Papain preparation (61.25 mg/l) Storage: 24 months at -20°C	- / -	100 ml	L 2223

Trypsin - EDTA

Trypsin/EDTA (0.05/0.02% in PBS without Ca/Mg) to passage monolayer cultures:

- Wash cell culture (monolayer) using physiological saline (w/o Ca/Mg).
- Apply a small quantity of Trypsin/EDTA with a pipette to cell culture flask; a thin film is sufficient.
- Incubate culture flask at +37°C for approx. 5 min.
- Centrifuge cell suspension, or transfer directly to a serum containing culture medium.
With higher trypsin concentrations, it is recommended to trypsinate at +2 - +8°C.
Formulation

Pack/Density

Unit

Cat.no.


(10x) Trypsin/EDTA (0,5% / 0,2%) in (10x) PBS without Ca²⁺, Mg²⁺ Storage: 36 months at -20°C	- / -	100 ml	L 2153



Trypsin (1:250) 0.25% in PBS without Ca²⁺, Mg²⁺ Storage: 36 months at -20°C	- / -	100 ml	L 2123



Trypsin (1:250) 2.5% in PBS without Ca²⁺, Mg²⁺ Storage: 36 months at -20°C	- / -	100 ml	L 2133



Trypsin/EDTA (0,05% / 0,02%) in PBS without Ca²⁺, Mg²⁺ Storage: 36 months at -20°C	- / -	100 ml	L 2143



Trypsin/EDTA (0,25% / 0,02%) in PBS without Ca²⁺, Mg²⁺ Storage: 36 months at -20°C	- / -	100 ml	L 2163

Tissue Desintegration Enzymes

Send mail to webmaster with questions or comments about this web site. Copyright © 2005 Gentaur BVBA Last modified: feb-07

Send mail to webmaster with questions or comments about this web site.
Copyright © 2005 Gentaur BVBA
Last modified: feb-07