Cell Dissociation - Cell Dissociation reagents

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.05% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4260.0100 (100mL) or 2 x C4260.0500 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.05% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4260.0100 (100mL) or 2 x C4260.0500 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.05% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4261.0100 (100mL) or 2 x C4261.0500 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.25% Trypsin is produced by dilution of Trypsin in HBSS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4288.0100 (100mL) or 2 x C4288.0500 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.25% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4267.0100 (100mL) or 2 x C4267.0500 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.25% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4259.0100 (100mL) or 2 x C4259.0500 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.25% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimum order size is 5 x C4262.0100 (100mL) or 2 x C4262.0500 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 0.5% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4261.0110 (100mL) or 2 x C4261.0510 (500mL).

Trypsin is a mixture of proteases isolated from porcine pancreas. 2.5% Trypsin is produced by dilution of Trypsin in PBS. Due to its protease activities, trypsin solutions are widely used for cell dissociation, routine cell culture passaging, or primary tissue dissociation. The trypsin concentration required for dissociation varies with cell type and experimental requirements. For this Genaxxon offers a variety of different Trypsin solutions with and without EDTA. Activity of the used Trypsin: 1g of Trypsin digests 250g of Casein substrate. Minimal order size: 5 x C4287.0100 (100mL) or 2 x C4287.0500 (500mL).

Accutase® as a cell detachment solution of proteolytic and collagenolytic enzymes useful for the routine detachment of cells from standard tissue culture plastic ware and adhesion coated plastic ware. The reagent is useful for creating single cell suspensions from clumped cell cultures for accurate cell counting, detachment of cells from primary tissue. It is much better suited for cell detachment than trypsin. Proven effective in detaching primary fibroblasts, endothelial cells, neurons, tumor cell lines, and insect cells. Accutase® does not contain mammalian or bacterial derived products. For that reason Accutase® virus or endotoxin contaminations are excluded. Once thawed Accutase® can be used for about 1 week if stored at +2°C to +8°C. Shelf-life at -20°C is 12 months. Note: Accutase is a registered trademark of Innovative Cell Technologies, Inc.

Clostridium histolyticum collagenase is an enzyme mixture of collagenase, clostripain and tryptic and proteolytic activities. Collagenase type I shows a balanced activity of collagenase, clostripain as well as tryptic and proteolytic activities. Type I Collagenase is recommended for cell preparation from epithelial, liver, lung tissue, tissue of the suprarenal gland and adipose tissue. The specific activity is >90 Mandl-Units per mg dry substance. Collagenase is produced by two separate and distinct genes in Clostridium histolyticum. Both genes have been cloned and sequenced (Yoshihara 1994). The colG gene codes for type I collagenase, a 936 amino acid protein, while the colH gene codes for type II collagenase, a 1021 amino acid protein. Both genes share 72% identity, the proteins only 43%. Both gene products can be present as two or more isoforms differing in molecular weight. Therefore collagenase mixtures can contain six to eight different proteins in a molecular weight range from 68 to 130 kDa. Substrate specificity studies have demonstrated that the colG gene prefers natural substrates such as intact collagen, compared to the colH gene product which preferentially digests short synthetic substrates (FALGPA) (Eckhard et al. 2009 and Matsushita 1999). General description:The treatment of tissue with collagenase causes the careful, selective degradation of the intercellular matrix, and does not affect the growth of the cells. The collagenase offered by Genaxxon bioscience is a mixture of different proteolytically active enzymes and requires calcium ions for both the catalytic activity and the binding to the collagen molecule. In contrast to vertebrate collagenase, Clostridium histolyticum collagenase digests native triple helix collagen into small peptides, which is the major use of collagenases in cell culture application. For optimal results, a well balanced mixture of proteolytic enzymes is necessary. Four different collagenases, Collagenase Type I >, Collagenase Type II >, Collagenase Type III > and Collagenase Type IV > are available for this. Type IV is usually used together with other enzymes, such as trypsin >, elastase or hyaluronidase >. Trypsin or Trypsin/EDTA > conventionally used in the cell culture degrades the matrix only slowly and can cause irreversible damage to the released cells. Therefore, we recommend Accutase > instead of trypsin or trypsin/EDTA. Clostridium collagenases belongs to the metalloproteases, a large family of proteases that shares a zinc-containing motif at the center of the active site (Gonzales and Robert-Baudouy 1996). The enzyme is reversibly inactivated at high pH values and irreversibly inactivated at low pH values. Inhibitors of collagenase include cysteine, EDTA, o-phenanthroline, 8-hydroxyquinoline-5-sulfonate, bipyridyl, 2,3-dimercaptopropanol or Hg2+, Pb2+, Cd2+, Cu2+. Collagenase is NOT inhibited by diisopropylphosphorofluoridate (DFP) or serum. Lyphilised Collagenase should be stored at +2°C to +8°C and remain stable without loss of activity for at least three years. Enzyme should be protected from moisture. Dissolved Collagenase can be aliquoted and stored at -20°C for one year. Conversion rates of collagenase activity units: 1 PZ U/mg ~ 3.9 FALGPA U/mg - 1 PZ U/mg ~ 1000 Mandl or CDU U/mg - 1 PZ U/mg ~ 10 HP U/mg (PZ-units according to Wünsch).

Clostridium histolyticum collagenase is an enzyme mixture of collagenase, clostripain and tryptic and proteolytic activities. Type II Collagenase is recommended for the preparation of cells from liver, bone, thyroid gland, heart and salivary gland and has a specific activity of >180 Mandl units per mg of dry matter. Collagenase is produced by two separate and distinct genes in Clostridium histolyticum. Both genes have been cloned and sequenced (Yoshihara 1994). The colG gene codes for type I collagenase, a 936 amino acid protein, while the colH gene codes for type II collagenase, a 1021 amino acid protein. Both genes share 72% identity, the proteins only 43%. Both gene products can be present as two or more isoforms differing in molecular weight. Therefore collagenase mixtures can contain six to eight different proteins in a molecular weight range from 68 to 130 kDa. Substrate specificity studies have demonstrated that the colG gene prefers natural substrates such as intact collagen, compared to the colH gene product which preferentially digests short synthetic substrates (FALGPA) (Eckhard et al. 2009 and Matsushita 1999). General description:The treatment of tissue with collagenase causes the careful, selective degradation of the intercellular matrix, and does not affect the growth of the cells. The collagenase offered by Genaxxon bioscience is a mixture of different proteolytically active enzymes and requires calcium ions for both the catalytic activity and the binding to the collagen molecule. In contrast to vertebrate collagenase, Clostridium histolyticum collagenase digests native triple helix collagen into small peptides, which is the major use of collagenases in cell culture application. For optimal results, a well balanced mixture of proteolytic enzymes is necessary. Four different collagenases, Collagenase Type I >, Collagenase Type II >, Collagenase Type III > and Collagenase Type IV > are available for this. Type IV is usually used together with other enzymes, such as trypsin >, elastase or hyaluronidase >. Trypsin or Trypsin/EDTA > conventionally used in the cell culture degrades the matrix only slowly and can cause irreversible damage to the released cells. Therefore, we recommend Accutase > instead of trypsin or trypsin/EDTA. Clostridium collagenases belongs to the metalloproteases, a large family of proteases that shares a zinc-containing motif at the center of the active site (Gonzales and Robert-Baudouy 1996). The enzyme is reversibly inactivated at high pH values and irreversibly inactivated at low pH values. Inhibitors of collagenase include cysteine, EDTA, o-phenanthroline, 8-hydroxyquinoline-5-sulfonate, bipyridyl, 2,3-dimercaptopropanol or Hg2+, Pb2+, Cd2+, Cu2+. Collagenase is NOT inhibited by diisopropylphosphorofluoridate (DFP) or serum. Lyphilised Collagenase should be stored at +2°C to +8°C and remain stable without loss of activity for at least three years. Enzyme should be protected from moisture. Dissolved Collagenase can be aliquoted and stored at -20°C for one year. Conversion rates of collagenase activity units: 1 PZ U/mg ~ 3.9 FALGPA U/mg - 1 PZ U/mg ~ 1000 Mandl or CDU U/mg - 1 PZ U/mg ~ 10 HP U/mg (PZ-units according to Wünsch).

Clostridium histolyticum collagenase is an enzyme mixture of collagenase, clostripain and tryptic and proteolytic activities. Collagenase type III shows normal Collagenase, but very low proteolytic activity. Type III Collagenase is recommended for preparation of cells from mamary gland and fetal cells and has a specific activity of 100 to 250 Mandl units per mg of dry matter. Collagenase is produced by two separate and distinct genes in Clostridium histolyticum. Both genes have been cloned and sequenced (Yoshihara 1994). The colG gene codes for type I collagenase, a 936 amino acid protein, while the colH gene codes for type II collagenase, a 1021 amino acid protein. Both genes share 72% identity, the proteins only 43%. Both gene products can be present as two or more isoforms differing in molecular weight. Therefore collagenase mixtures can contain six to eight different proteins in a molecular weight range from 68 to 130 kDa. Substrate specificity studies have demonstrated that the colG gene prefers natural substrates such as intact collagen, compared to the colH gene product which preferentially digests short synthetic substrates (FALGPA) (Eckhard et al. 2009 and Matsushita 1999). General description:The treatment of tissue with collagenase causes the careful, selective degradation of the intercellular matrix, and does not affect the growth of the cells. The collagenase offered by Genaxxon bioscience is a mixture of different proteolytically active enzymes and requires calcium ions for both the catalytic activity and the binding to the collagen molecule. In contrast to vertebrate collagenase, Clostridium histolyticum collagenase digests native triple helix collagen into small peptides, which is the major use of collagenases in cell culture application. For optimal results, a well balanced mixture of proteolytic enzymes is necessary. Four different collagenases, Collagenase Type I >, Collagenase Type II >, Collagenase Type III > and Collagenase Type IV > are available for this. Type IV is usually used together with other enzymes, such as trypsin >, elastase or hyaluronidase >. Trypsin or Trypsin/EDTA > conventionally used in the cell culture degrades the matrix only slowly and can cause irreversible damage to the released cells. Therefore, we recommend Accutase > instead of trypsin or trypsin/EDTA. Clostridium collagenases belongs to the metalloproteases, a large family of proteases that shares a zinc-containing motif at the center of the active site (Gonzales and Robert-Baudouy 1996). The enzyme is reversibly inactivated at high pH values and irreversibly inactivated at low pH values. Inhibitors of collagenase include cysteine, EDTA, o-phenanthroline, 8-hydroxyquinoline-5-sulfonate, bipyridyl, 2,3-dimercaptopropanol or Hg2+, Pb2+, Cd2+, Cu2+. Collagenase is NOT inhibited by diisopropylphosphorofluoridate (DFP) or serum. Lyphilised Collagenase should be stored at +2°C to +8°C and remain stable without loss of activity for at least three years. Enzyme should be protected from moisture. Dissolved Collagenase can be aliquoted and stored at -20°C for one year. Conversion rates of collagenase activity units: 1 PZ U/mg ~ 3.9 FALGPA U/mg - 1 PZ U/mg ~ 1000 Mandl or CDU U/mg - 1 PZ U/mg ~ 10 HP U/mg (PZ-units according to Wünsch).

Clostridium histolyticum collagenase is an enzyme mixture of collagenase, clostripain and tryptic and proteolytic activities. Collagenase type IV has low tryptic, high collagenase and normal clostripain activity. Type I Collagenase is recommended for cell preparation from epithelial, liver, lung tissue, tissue of the suprarenal gland and adipose tissue. The specific activity of >900 Mandl units per mg of dry matter. Collagenase is produced by two separate and distinct genes in Clostridium histolyticum. Both genes have been cloned and sequenced (Yoshihara 1994). The colG gene codes for type I collagenase, a 936 amino acid protein, while the colH gene codes for type II collagenase, a 1021 amino acid protein. Both genes share 72% identity, the proteins only 43%. Both gene products can be present as two or more isoforms differing in molecular weight. Therefore collagenase mixtures can contain six to eight different proteins in a molecular weight range from 68 to 130 kDa. Substrate specificity studies have demonstrated that the colG gene prefers natural substrates such as intact collagen, compared to the colH gene product which preferentially digests short synthetic substrates (FALGPA) (Eckhard et al. 2009 and Matsushita 1999). General description:The treatment of tissue with collagenase causes the careful, selective degradation of the intercellular matrix, and does not affect the growth of the cells. The collagenase offered by Genaxxon bioscience is a mixture of different proteolytically active enzymes and requires calcium ions for both the catalytic activity and the binding to the collagen molecule. In contrast to vertebrate collagenase, Clostridium histolyticum collagenase digests native triple helix collagen into small peptides, which is the major use of collagenases in cell culture application. For optimal results, a well balanced mixture of proteolytic enzymes is necessary. Four different collagenases, Collagenase Type I >, Collagenase Type II >, Collagenase Type III > and Collagenase Type IV > are available for this. Type IV is usually used together with other enzymes, such as trypsin >, elastase or hyaluronidase >. Trypsin or Trypsin/EDTA > conventionally used in the cell culture degrades the matrix only slowly and can cause irreversible damage to the released cells. Therefore, we recommend Accutase > instead of trypsin or trypsin/EDTA. Clostridium collagenases belongs to the metalloproteases, a large family of proteases that shares a zinc-containing motif at the center of the active site (Gonzales and Robert-Baudouy 1996). The enzyme is reversibly inactivated at high pH values and irreversibly inactivated at low pH values. Inhibitors of collagenase include cysteine, EDTA, o-phenanthroline, 8-hydroxyquinoline-5-sulfonate, bipyridyl, 2,3-dimercaptopropanol or Hg2+, Pb2+, Cd2+, Cu2+. Collagenase is NOT inhibited by diisopropylphosphorofluoridate (DFP) or serum. Lyphilised Collagenase should be stored at +2°C to +8°C and remain stable without loss of activity for at least three years. Enzyme should be protected from moisture. Dissolved Collagenase can be aliquoted and stored at -20°C for one year. Conversion rates of collagenase activity units: 1 PZ U/mg ~ 3.9 FALGPA U/mg - 1 PZ U/mg ~ 1000 Mandl or CDU U/mg - 1 PZ U/mg ~ 10 HP U/mg (PZ-units according to Wünsch).

1% EDTA solution in PBS without Calcium and Magnesium. A 1% solution is a 5-time concentrated solution. Normally a 0.05% EDTA solution in PBS is used for the following applications: - disengage attached and clumped cells. - in procedures generating dendritic cells from monocytes.- PBS+EDTA to get better cell sorting. - PBS/EDTA may also be used directly without addition of trypsine for very gentle cell dissociation.

Trypsin from bovine pancreas, Tryptic activity (BAEE): min. 2500 U/mg. Lyophilised. No P. aeruginosa, Salmonella, Staphyloccus aureus detectable. Activity: 1g of Trypsin digests 250g of Casein substrate. Synonym: Peptidylpeptid-Hydrolase (E.C. 3.4.21.4).

Trypsin 1:250 from porcine pancreas (240 - 260 USP U/mg). Contains Chymotrypsin, Elastase and other non proteolytic activities. The native form of Trypsin consists of a single chain polypeptide of 223 amino acid residues, produced by the removal of the N-terminal hexapeptide from trypsinogen which is cleaved at the Lys - lle peptide bond. The sequence of amino acids is cross-linked by 6 disulfide bridges. Trypsin is a member of the serine protease family. ApplicationFor trypsin digestion of peptides, use a ratio of about 1:100 to 1:20 for trypsin:peptide. The typical use for this product is in removing adherent cells from a culture surface. The concentration of trypsin necessary to dislodge cells from their substrate is dependent primarily on the cell type and the age of the culture. Trypsins have also been used for the re-suspension of cells during cell culture, in proteomics research for digestion of proteins and in various in-gel digestionsns. Additional applications include assessing crystallization by membrane-based techniques and in a study to determine that protein folding rates and yields can be limited by the presence of kinetic traps. Serine protease inhibitors, including DFP, TLCK, APMSF, AEBSEF, and aprotinin, amongst others, will inhibit Trypsin. Preparation of Trypsin solutionsThis product is a lyophilized powder which can be easily dissolved in aqueous solutions, e.g. in Hank′s Balanced Salt Solution at 25mg/mL. For applications that require EDTA, solubilizing trypsin should be done with a buffered salt solution contaiing no Ca2+ or Mg2+. Caution: Solutions in 1 mM HCl are stable for 1 year in aliquots and stored at -20°C. The presence of Ca2+ will also diminish the self-autolysis of trypsin and maintain its stability in solution. Trypsin will also retain most of its activity in 2.0 M urea, 2.0 M guanidine HCl, or 0.1% (w/v) SDS. Unit Definition: One BAEE unit will produce a ΔA253 of 0.001 per min at pH 7.6 at 25° C using BAEE as substrate. One BTEE unit = 320 ATEE units. Reaction volume = 3.2 mL (1 cm light path).

Zymolyase®, produced by a submerged culture of Arthrobacter luteus, has strong lytic activity against living yeast cell walls to produce protoplast or spheroplast of various strains of yeast cells. An essential enzyme for the lytic activity of Zymolyase® is β-1,3-glucan laminaripentaohydrolase. It hydrolyzes linear glucose polymers with β-1,3-linkages and releases specifically laminaripentaose as the main and minimum product unit. Reported to be useful for lysis of Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloeckera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Torulopsis, Saccharomyces, Saccharomycopsis, Saccharomycodes, and Schwanniomyces species. There are two preparations of Zymolyase®, Zymolyase®-20T (C4352) and Zymolyase®-100T (C4366), having lytic activity of 20,000 units/g and 100,000 units/g respectively. The activity was tested towards brewer’s yeast cells (Saccharomyces cerevisiae, resting stage) or towar yeast cells of Saccharomyces uvarum (IFO 0565) cultured statically in malt extract medium. Zymolyase®-20T is ammonium sulfate precipitate while Zymolyase®-100T is a further purified preparation by affinity chromatography. Lytic activity varies depending on yeast strain, growth stage of yeast, or cultural conditions. Further informations related to Zymolyase® can be obtained from the data sheet and the references given there.

Zymolyase®, produced by a submerged culture of Arthrobacter luteus, has strong lytic activity against living yeast cell walls to produce protoplast or spheroplast of various strains of yeast cells. An essential enzyme for the lytic activity of Zymolyase® is β-1,3-glucan laminaripentaohydrolase. It hydrolyzes linear glucose polymers with β-1,3-linkages and releases specifically laminaripentaose as the main and minimum product unit. Reported to be useful for lysis of Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloeckera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Torulopsis, Saccharomyces, Saccharomycopsis, Saccharomycodes, and Schwanniomyces species. There are two preparations of Zymolyase®, Zymolyase®-20T (C4352) and Zymolyase®-100T (C4366) , having lytic activity of 20,000 units/g and 100,000 units/g respectively. The activity was tested towards brewer’s yeast cells (Saccharomyces cerevisiae, resting stage) or towar yeast cells of Saccharomyces uvarum (IFO 0565) cultured statically in malt extract medium. Zymolyase®-20T is ammonium sulfate precipitate while Zymolyase®-100T is a further purified preparation by affinity chromatography. Lytic activity varies depending on yeast strain, growth stage of yeast, or cultural conditions. Further informations related to Zymolyase® can be obtained from the data sheet and the references given there.