Tissue Dissociation is used to isolate cells from tissues. The aim is to get the highest number of cells to the best possible quality for further experiments. Critical issue is the quality of used enzymes, purified or crude.
In many labs crude is used which has the big disadvantage of enzymatic contamination. This leads to a low reproducibility and the need to test every new batch for your application.
To overcome these disadvantages we offer only highly purified enzyme systems and guarantee that - once tested - these work every time in the same way. Results will be reproducible and further batch testing is redundant.
Collagenases are enzymes that digest collagen the major fibrous component of animal and human extracellular connective tissue. These enzymes are prepared from C. histolyticum and are often used as crude enzyme preparation. The advantage of bacterial endopeptidases is that these enzymes will digest the insoluble collagen as well as soluble collagen fragments. The disadvantage of using crude collagenase preparations are the contaminating foreign enzyme activities (tryptic, caseinase and clostripain) which prevents reproducibility. Running out of collagenase scientists have to test new batches to find the right one for their application and tissue. Avoiding this batch testing procedure repeatedly would be a big step forward and improve productivity. Highly purified enzyme preparations will make this lot procedure superfluous.
Our partner VitaCyte offers only highly purified enzymes (collagenase and protease) as pure preparation and enzymatic blends. Recently we launched the DE collagenase product family which is characterized by increasing collagenase activities with the same amount of protease activity.
DE Collagenase consists of increasing amounts of enriched collagenase (> 85% pure) with minimal protease activity add to fixed amount of purified BP Protease. The enriched collagenase has high collagen degradation activity and very low contamination with protease (<0.5%). The following blends are available:
Thermolysin a neutral protease prepared from Bacillus thermoproteolyticus is a thermostable metalloproteinase used for isolation of islets from rodent, canine, monkey and human pancreas.
Clostripain is an endoproteinase Arg-c prepared from B, clostridium histolyticum and cleaves proteins on the carboxyl peptide bond of arginin. Clostripain is used as a supplement to collagenase in tissue dissociation applications often along with Thermolysin or BP Protease. Each tissue type and primary cell of interest has its own clostripain requirement.
BP Protease (Neutral Protease, Dispase) is prepared from Bacillus polymyxa produced without components of animal orgin. BP Protease was designed to isolate islets, hepatocytes and adipose derived stromal cells (ADSC).
Enzymatic blends are mixtures of purified collagenase and purified proteases designed for special applications to increase reproducibility and lot-to-lot consistency.
We offer tested and pretested blends for rodent islets, adipose derived stem cells and human hepatocytes