3. Collagenase I, 100 mg $180

Description

Product Description

Collagenase is a proteolytic enzyme commonly used for digestion of extracellular matrix proteins. As a peptidase, collagenase specifically recognizes the Pro-X-Gly-Pro sequence (which is highly frequent in collagen but rare in other proteins) and cleaves the peptide bond between the amino acid (X) and glycine (Gly). Commercially available collagenase is derived from Clostridium histolyticum, a type of bacteria, and it is obtained as a crude enzyme preparation. There are different types of collagenase, including Type I collagenase, Type II collagenase, Type III collagenase, Type IV collagenase, and Type V collagenase, which vary in their composition and degradation characteristics, leading to different applications.

Type I collagenase contains a relatively balanced mixture of various enzyme activities, including collagenase, caseinase, clostripain, and trypsin, and it is typically used for the preparation of primary cells from epithelial tissues, liver, lung, adipose tissue, and adrenal tissue.

Type II collagenase has higher clostripain activity and is commonly used for the preparation of primary cells from heart, bone, liver, thymus, salivary glands, and cartilage.

Type III collagenase has lower secondary protease activity and is frequently used for the preparation of primary cells from mammary tissue.

Type IV collagenase has lower trypsin activity and is usually used for the preparation of islet cells and myeloid cells.

Type V collagenase is a partially purified form with higher collagenase and caseinase activity and lower trypsin levels, suitable for the preparation of islets.

Please note that the provided translation is a general description of the different types of collagenase and their applications. The specific characteristics and applications may vary depending on the commercial product or supplier.

Basic Properties:

English Name: Collagenase I Aliases: Clostridiopeptidase A CAS: 9001-12-1 Enzymatic Activity: ≥125 CDU/mg, solid Appearance: Brown or tan crystalline powder Storage Conditions: -20°C Unit: Bottle Specification: 100mg/bottle MDL Number: MFCD00130830 EC Number: 232-582-9 Related Categories: Protein Derivatives Solubility: Soluble in water Shelf Life: 24 months

CDU Definition: One casein digestion unit (CDU) is defined as the amount of enzyme required to liberate 1 μmol of tyrosine from collagen protein from bovine Achilles tendon under the specified conditions of 37°C, pH 7.4, and the presence of calcium ions.

Storage and Transportation: Transport with dry ice and store at -20°C.

Instructions for Use:

The usage and dosage of biochemical reagents should be determined based on the customer’s experimental purposes and existing literature or experimental methods in books. The following instructions are provided for reference only.

1. Preparation of Collagenase Storage Solution:

Add 100µL of HBSS (Hank’s balanced salt solution) containing Ca2+ and Mg2+ to each vial of 100mg collagenase. Gently vortex and shake to dissolve completely, preparing a storage solution with a concentration of 1g/ml (or 1000×). Then, filter the solution through a low-protein-binding 0.22μm membrane to sterilize and divide into smaller aliquots. Store the aliquots at -20°C, protected from light.

Thaw the solution on ice before use, avoiding repeated freeze-thaw cycles. The commonly used concentrations for tissue and cell dispersion are 0.5-2.5mg/ml, while for cartilage digestion, the commonly used concentration is 1-2mg/ml. The optimal working concentration should be determined based on specific experimental conditions or by referring to relevant literature.

2. Tissue Dissociation:

1. Use a sterile surgical knife or scissors to cut the tissue into pieces of 3-4mm in size.
2. Wash the tissue pieces several times with HBSS containing Ca2+ and Mg2+.
3. Add an adequate amount of HBSS containing Ca2+ and Mg2+ to submerge the tissue pieces, and add collagenase to the desired working concentration.
4. Incubate at 37°C for 4-18 hours. The use of a horizontal shaker and supplementation with 3mM CaCl2 during digestion can improve digestion efficiency.
5. Dispersed cells can be collected using stainless steel or nylon mesh filters and kept for further use. Undissociated tissues can be incubated with an additional appropriate amount of fresh collagenase working solution at 37°C.
6. Wash the collected cells several times with collagenase-free HBSS.
7. Resuspend the cells in cell culture medium and use an automated cell counter or other methods to determine the density of viable cells.
8. Inoculate the cells onto cell culture dishes using suitable cell culture medium.

Precautions:

For your safety and health, please wear laboratory attire and disposable gloves during the operation.

Safety Information:

Warning Statements: P261, P305+P351+P338, P342+P311

WGK: 1

Note: Product improvements may occur. Please refer to the actual label information.

This product is for research purposes only and is not intended for clinical diagnosis.