Trypsin 0.25% Solution (1X)
Trypsin solutions are enzyme preparations derived from porcine pancreas, generally used for the isolation and dissociation of anchorage-dependent mammalian cells and tissues. The concentration of trypsin necessary for the successful isolation of cells depends primarily on the cell type and the age of the culture.

Trypsin 0.25% is the most commonly used trypsin solution. It contains 2.5 gm/L of trypsin (1:250) in Hanks’ Balanced Salt Solution without calcium and magnesium salts.

Trypsin 2.5% Solution (10X)
Trypsin solutions are enzyme preparations derived from porcine pancreas, generally used for the isolation and dissociation of anchorage-dependent mammalian cells and tissues. The concentration of trypsin necessary for the successful isolation of cells depends primarily on the cell type and the age of the culture.

Trypsin 2.5% is a 10X concentrate of Trypsin 0.25%, the most commonly used trypsin solution. It contains 25 gm/L of trypsin (1:250) in normal saline solution and must be diluted in an appropriate buffer before use.

Trypsin 0.25%-EDTA 1mM Solution (1X)
Trypsin solutions are enzyme preparations derived from porcine pancreas, generally used for the isolation and dissociation of anchorage-dependent mammalian cells and tissues. The concentration of trypsin necessary for the successful isolation of cells depends primarily on the cell type and the age of the culture.

Trypsin 0.25% is the most commonly used trypsin solution. EDTA (Ethylenediaminetetraacetic acid), a chelating agent that neutralizes calcium and magnesium ions, is added to increase the enzymatic activity on the cells and to facilitate release of individual cells from the culture vessel. Calcium and magnesium ions enhance cell-to-cell adhesion and obscure the peptide bonds on which trypsin acts. This trypsin solution contains 2.5 gm/L of trypsin (1:250) and 0.38 gm/L EDTA•4Na in Hanks’ Balanced Salt Solution without calcium and magnesium salts.

Trypsin 0.05%-EDTA 0.53mM Solution (1X)
Trypsin solutions are enzyme preparations derived from porcine pancreas, generally used for the isolation and dissociation of anchorage-dependent mammalian cells and tissues. The concentration of trypsin necessary for the successful isolation of cells depends primarily on the cell type and the age of the culture.

Trypsin 0.05% is the trypsin solution of choice for more sensitive cell lines. EDTA (Ethylenediaminetetraacetic acid), a chelating agent that neutralizes calcium and magnesium ions, is added to increase the enzymatic activity on the cells and to facilitate release of individual cells from the culture vessel. Calcium and magnesium ions enhance cell-to-cell adhesion and obscure the peptide bonds on which trypsin acts. This trypsin solution contains 0.5 gm/L of trypsin (1:250) and 0.2 gm/L EDTA•4Na in Hanks’ Balanced Salt Solution without calcium and magnesium salts.

Trypsin 0.5%-EDTA 5.3mM Solution (10X)
Trypsin is an enzyme preparation derived from porcine pancreas, generally used for the isolation and dissociation of anchorage-dependent mammalian cells and tissues. The concentration of trypsin necessary for the successful isolation of cells depends primarily on the cell type and the age of the culture.

Trypsin 0.5% is a 10X concentrate of Trypsin 0.05%, the trypsin solution of choice for more sensitive cell lines. EDTA (Ethylenediaminetetraacetic acid), a chelating agent that neutralizes calcium and magnesium ions, is added to increase the enzymatic activity on the cells and to facilitate release of individual cells from the culture vessel. Calcium and magnesium ions enhance cell-to-cell adhesion and obscure the peptide bonds on which trypsin acts. This trypsin solution contains 5.0 gm/L of trypsin (1:250) and 2.0 gm/L EDTA•4Na in normal saline solution and must be diluted in an appropriate buffer before use.

Trypsin, 1X crystallized
Trypsin is a pancreatic serine protease with substrate specificity based upon positively charged lysine and arginine side chains. The molecular weight of trypsin is 23,800 dalton and the optimum pH for enzymatic activity is 8.0.

Source: bovine pancreas
1X crystallized. Dialyzed against 1mM HCl and lyophilized.
Activity: ≥150 units per mg protein (≥8,625 BAEE/2,875 USP/NF units per mg protein)

Trypsin, 2X crystallized
Trypsin is a pancreatic serine protease with substrate specificity based upon positively charged lysine and arginine side chains. The molecular weight of trypsin is 23,800 dalton and the optimum pH for enzymatic activity is 8.0.

Source: bovine pancreas
2X crystallized. Dialyzed against 1mM HCl and lyophilized.
Activity: ≥180 units per mg protein (≥10,350 BAEE/3,450 USP/NF units per mg protein)

Trypsin Inhibitor
Cell isolation procedures occasionally call for a trypsin inhibitor, usually the inhibitor from soybean. The trypsin inhibitor from soybean inactivates trypsin on an equimolar basis and to a lesser extent chymotrypsin; however, it exhibits no effects on the esterolytic, proteolytic or elastolytic activities of porcine elastase. Its molecular weight is 21,500 ± 800 daltons and the optimum pH for enzymatic activity is 7.0.

Source: soybean
A partially purified acetone powder.
Activity: 1mg inhibits ≥0.75 mg trypsin

Versene 1:5000 Solution (1X)
Versene 1:5000 is a 1X solution containing 0.2 gm/L EDTA•4Na in phosphate–buffered saline.
Concentration: 0.53mM EDTA•4Na

EDTA is a calcium chelator used to eliminate the inhibition of enzyme catalyzed reactions due to traces of heavy metals.

Ethylenediaminetetraacetic Acid (EDTA)
EDTA tetrasodium salt dihydrate (C10H12N2Na4O8•2H2O), powder
EDTA is a calcium chelator used to eliminate the inhibition of enzyme catalyzed reactions due to traces of heavy metals.

Collagenase, Type 1
Collagenases are widely used in enzymatic primary cell isolation and tissue dissociation procedures. Collagenase, Type 1 is a crude collagenase preparation containing average amounts of assayed activities (collagenase, caseinase, clostripain, and tryptic activities). It is generally recommended for epithelial, liver, lung, fat, and adrenal tissue cell preparations.

Source: Clostridium histolyticum
A dialyzed, lyophilized powder
Activity: ≥125 units per mg dry weight
One unit releases one µmole of L-leucine equivalents from collagen in 5 hours at 37°C, pH 7.5.

Collagenase, Type 2
Collagenases are widely used in enzymatic primary cell isolation and tissue dissociation procedures. Collagenase, Type 2 is a crude collagenase preparation containing greater clostripain activity. It is generally used for heart, bone, muscle, thyroid and cartilage tissue cell preparations.

Source: Clostridium histolyticum
A dialyzed, lyophilized powder
Activity: ≥125 units per mg dry weight
One unit releases one µmole of L-leucine equivalents from collagen in 5 hours at 37°C, pH 7.5.

Collagenase, Type 3
Collagenases are widely used in enzymatic primary cell isolation and tissue dissociation procedures. Collagenase, Type 3 is a crude collagenase preparation that is usually selected because of its low proteolytic activity. It is commonly used for primary mammary cell isolation.

Source: Clostridium histolyticum
A dialyzed, lyophilized powder
Activity: ≥100 units per mg dry weight
One unit releases one µmole of L-leucine equivalents from collagen in 5 hours at 37°C, pH 7.5.

Collagenase, Type 4
Collagenases are widely used in enzymatic primary cell isolation and tissue dissociation procedures. Collagenase, Type 4 is a crude collagenase preparation that is prepared to contain lower tryptic activity levels to limit damage to cell membrane proteins and receptors but with normal to above normal collagenase activity. It is commonly used for pancreatic islet cell isolation and other applications where receptor integrity is crucial.

Source: Clostridium histolyticum
A dialyzed, lyophilized powder
Activity: ≥160 units per mg dry weight
One unit releases one µmole of L-leucine equivalents from collagen in 5 hours at 37°C, pH 7.5.

Collagenase, Purified
This chromatographically purified Collagenase preparation is widely used in enzymatic primary cell isolation and tissue dissociation procedures, usually in combination with secondary enzymes such as elastase, hyaluronidase, etc. It is composed of two separable but very similar collagenases. It contains ≤50 caseinase units per milligram.

Source: Clostridium histolyticum
A lyophilized powder
Activity: ≥500 units per mg dry weight
One unit releases one µmole of L-leucine equivalents from collagen in 5 hours at 37°C, pH 7.5.

Chymotrypsin, Alpha
Chymotrypsin is a protease that preferentially catalyzes the hydrolysis of peptide bonds involving the aromatic amino acids tyrosine, phenylalanine, and tryptophan. It also acts upon the peptide bonds of leucyl, methionyl, asparagenyl and glutamyl residues, and the amides and esters of susceptible amino acids.

Chymotrypsin is used to a limited extent in tissue dissociation, usually in combination with trypsin and elastase.

Source: Bovine pancreas
3X crystallized alpha chymotrypsin (activation products of 3X crystallized zymogen)
Dialyzed against 1mM HCl and lyophilized
Activity: ≥45 units per mg protein
One unit hydrolyzes 1 µmole of BTEE per minute at 25°C, pH 7.8 in the presence of calcium.

Deoxyribonuclease I
Often as a result of cell damage, deoxyribonucleic acid (DNA) leaks into the dissociation medium increasing viscosity and causing handling problems. Purified deoxyribonuclease is sometimes included in cell isolation procedures to digest the DNA without damaging the intact cells.

Bovine pancreatic deoxyribonuclease is an endonuclease that splits phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide yielding polynucleotides with a free hydroxyl group at the 3’ position and a phosphate group at the 5’ position.

Source: Bovine pancreas
Purified precrystalline DNase. A lyophilized powder.
Activity: ≥2,000 Kunitz units per mg dry weight
0.005 Kunitz unit digests 1µg of λDNA in 10 minutes at 37°C in 50mM Tris, 1mM Mg²+, pH 7.8 in a 50 µl reaction.

Dispase (Neutral Protease)
Dispase (Neutral Protease) is a bacterial enzyme produced by Bacillus polymyxa that hydrolyses N-terminal peptide bonds of non-polar amino acid residues and is classified as an amino-endopeptidase. Its mild proteolytic action makes the enzyme especially useful for the isolation of primary and secondary cell cultures, since it maintains cell membrane integrity. Dispase is also frequently used as a secondary enzyme in conjunction with collagenase and/or other proteases in many primary cell isolation and tissue dissociation applications. Since Dispase dissociates fibroblast-like cells more efficiently than epithelial-like cells it is also used for differential cell isolation and cell culture applications. Dispase is inhibited by EDTA.

Source: Bacillus polymyxa
Chromatographically purified. A lyophilized powder.
Activity: ≥6 units per mg dry weight
One unit releases Folin positive amino acids equivalent to 1µmole tyrosine per minute from casein at 37°C, pH 7.5.

Elastase
Pancreatic Elastase is a serine protease with specificity for peptide bonds adjacent to neutral amino acids. It also exhibits esterase and amidase activity. While Elastase will hydrolyze a wide variety of protein substrates, it is unique among proteases in its ability to hydrolyze native elastin, a substrate not attacked by trypsin, chymotrypsin or pepsin. It is produced in the pancreas as an inactive zymogen, proelastase, and activated in the duodenum by trypsin. Elastase is also found in blood components and bacteria.

Elastin is found in highest concentration in the elastic fibers of connective tissues; therefore, elastase is frequently used to dissociate tissues that contain a large amount of connective tissue. For this purpose, it is usually used with other enzymes such as collagenase, trypsin, and chymotrypsin. Elastase is the enzyme of choice for the isolation of Type II cells from the lung.

Elastase has a molecular weight of 25,900 dalton, and an optimum pH for enzymatic activity of 8.5. It is unstable at pH ≤3.5.

Source: Porcine pancreas
2X crystallized; dialyzed and lyophilized
Activity: ≥3 units per mg protein
One unit cleaves 1µmole of SucAla3NA per minute at 25°C, pH 8.0.

Hyaluronidase
Hyaluronidase is a polysaccharidase with specificity for endo-N-acetylhexosaminic bonds between
2-acetoamido-2-deoxy-beta-D-glucose and D-glucuronate. These bonds are common in hyaluronic acid and chondroitin sulfate A and C. Hyaluronidase is often used for the dissociation of tissues, usually in combination with a crude protease such as collagenase, since the aforementioned substances are found in high concentrations in the ground substance of virtually all connective tissues. The optimum pH range is 4.5 to 6.0.

Source: Bovine testes
A partially purified, dialyzed, lyophilized powder
Activity: ≥300 USP/NF units per mg dry weight
One unit corresponds to a USP/NF Unit and is referenced to a standard NF Hyaluronidase.

Papain
Papain is a sulfhydryl protease from Carica papaya latex. It has a wide specificity and will degrade most protein substrates more extensively than the pancreatic proteases. Papain breaks down the intercellular matrix of cartilage. It also exhibits esterase activity. With some tissues Papain has proved less damaging and more effective that other proteases.

Papain is activated by cystein, sulfide, and sulfite. Stabilizing agents are EDTA, cysteine and dimercaptoethanol.

Papain has a molecular weight of 23,000 daltons and an optimum pH range of 6.0-7.0.

Source: Carica papaya latex
2X crystallized; lyophilized powder
Activation prior to use is recommended.
Activity: ≥15 units per mg protein
One unit hydrolyzes 1 µmole of BAEE per minute at 25°C, pH 6.2, after activation in a solution containing 1.1mM EDTA, 0.067mM ß-mercaptoethanol and 5.5mM cysteine-HCl for 30 minutes.

Pepsin A
Pepsin is an acid protease. Its inactive zymogen precursor, pepsinogen, is produced in the stomach mucosa. There are several pepsins designated A, B, C, and D. Pepsin A, the major component, has a molecular weight of 35,000 daltons and an optimum pH for enzymatic activity of approximately 1.0 for substrates such as casein or hemoglobin if the substrate is native protein. Pepsin cleaves proteins preferentially at the carboxylic groups of aromatic amino acids such as phenylalanine and tyrosine. It will not cleave at bonds containing valine, alanine, or glycine. Pepsinogen activates to pepsin below pH 5. Pepsin is unstable above pH 6.

Source: Porcine stomach
2X crystallized from dilute alcohol; lyophilized powder
Activity: ≥2,500 units per mg dry weight
One unit releases 0.001 A280 as TCA soluble hydrolysis products per minute at 37°C using denatured hemoglobin substrate.