How to prepare Dispase II?

Wash the tissue fragments in sterile PBS. Incubate the fragments in the Dispase® solution (2.4 U/ml – 0.6 U/ml) at 37°C. Stir slowly at 37°C until the tissue is sufficiently dissolved. total reaction time by counting the cells.

How do you dilute Dispase?

Prepare the Dispase Solution: Dissolve 2mg of Dispase into 1 ml of DMEM/F12. 9. Alternatively, make a 10x stock of Dispase (20mg/ml) and store in 1 ml aliquots in the -20C freezer. These aliquots can be thawed and diluted with 9mls of DMEM/F12.

What is dispase used for?

Dispase is a protease that is suitable for the gentle dissociation of a wide variety of tissues. Incubation of minced tissue with pre-warmed dispase and gentle agitation will liberate cells with minimal cell damage. Pre-warmed dispase can also be used to harvest cells from tissue culture plastic.

What is the 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. They are considered a virulence factor, facilitating the spread of gas gangrene.

How do you use Dispase?

I prefer to use 5 times the volume of dispase used per wash. Spin at 200xg for 5 minutes at room temperature. Alternatively, you can just let the clusters settle with gravity for 5 minutes in the hood. This might help to eliminate single cells that are thought to contribute to karyotype problems in hPSC cultures.

What is Dispase II?

Dispase II (neutral protease) is an amino-endo peptidase that hydrolyzes the N-terminal peptide bonds of non-polar amino acid residues. This sequence is found with high frequency in collagen. Dispase II has a mild proteolytic action that makes it useful for the isolation and routine passaging of primary cells.

Is collagenase regulated?

These levels can be regulated by parathyroid hormone (PTH), retinoic acid, and insulin-like growth factors, as well as the degree of matrix mineralization. Thus, there are many levels of regulation of collagenase in bone perhaps constraining what would otherwise be a rampant enzyme.

What is the function of Dispase?