What is function of collagenase?
Collagenases are enzymes that break the peptide bonds in collagen. They assist in destroying extracellular structures in the pathogenesis of bacteria such as Clostridium.
Where does collagenase cleave?
cartilage
Collagenase is the only proteinase that can cleave the triple helix of type II collagen in cartilage and type I and III collagens in other tissues.
Does collagenase need calcium?
Activators/Cofactors: Collagenase activity is stabilized by 0.1 mole calcium ions (Ca2+) per mole of enzyme. 4 Calcium ions also facilitate binding to the collagen molecule. 18 Zinc ions (Zn2+) are required for activity, but are tightly bound to the collagenase during purification.
How do you reconstitute collagenase?
Reconstitute the lyophilized collagenase powder (50 mg) in phosphate buffered saline or other pH neutral medium. A typical concentration of 1 mg/ml may be used.
What is Type 4 collagenase?
Collagenase is a protease consisting of a single polypeptide chain approximately 1,000 amino acid residues in length. Collagenase Type IV contains low levels of tryptic activity and is frequently used for pancreatic islets (Taguchi et al.) and for applications where maintenance of receptor integrity is required.
How many types of collagenase are there?
Rockland provides 4 types of collagenases, in different preparations, with specific affinities for use in different Tissue types.
Is collagenase a metalloproteinase?
The 72-kDa gelatinase/type IV collagenase (MMP-2) is a member of the matrix metalloproteinase (MMP) family of enzymes. This enzyme is known to cleave type IV collagen as well as degrade denatured collagens.
How do you dissolve Collagenase?
Add 1 mL Hanks’ Balanced Salt Solution (HBSS) with calcium and magnesium directly to 1 g vial of Collagenase. Vortex gently to ensure complete dissolution. Transfer to a clean tube.
How do you dilute IV collagenase?
Preparation of Collagenase Solution: Dissolve Collagenase Type IV in DMEM/F-12 medium to a concentration of 1 mg/ml and filter sterilize with a 0.22 µm pore size filter unit.
What is the difference between collagenase types?
Type 1 partially purified collagenase has the original balance of collagenase, caseinase, clostripain and tryptic activities. Type 2 contains higher relative levels of protease activity, particularly clostripain. Type 5 contains higher collagenase and caseinase values.
What is collagenase IV?
Collagenase IV is an enzyme purified from Clostridium histolyticum intended for cell and tissue disaggregation of islet tissues/cells (insulin receptor sites). Collagenase is unique among proteases in its ability to degrade the triplehelical native collagen fibrils commonly found in connective tissue.
How much collagenase should I use?
When this enzyme is tested for suitability for the release of hepatocytes, the collagenase is used at approximately 1 mg/ml in a total volume of 100 ml for each rat liver. If excessive cell death is observed with concentrations used with previous lots, the new lot used might have a higher specific activity.
