| Description | Collagenase NB 5 is a crude collagenase that contains collagenolytic and additional enzymatic activities including clostripain and neutral protease. The balanced ratio of these activities ensures gentle and efficient tissue dissociation.Collagenase NB 5 is applicable to tissue dissociation such as Collagenase NB 5 is a crude collagenase that contains collagenolytic and additional enzymatic activities including clostripain and neutral protease. The balanced ratio of these activities ensures gentle and efficient tissue dissociation.Collagenase NB 5 is applicable to tissue dissociation such as isolation of human and rodent hepatocytes, chondrocytes, neuronal and endothelial cells, fibroblasts, cells from adipose tissue (e.g. ADSCs), stem cells and rodent osteoblasts.ApplicationCollagenase NB 5 is suitable for dissociation of a broad variety of tissue types. SpecificationsContains class I and class II collagenase (1) and a balanced ratio of proteolytic side activities.Activity (Wünsch): ≥ 0.10 U/mgMicrobiological examination: Sterility, absence of ClostridiaSterile according to Ph. Eur.(Clostridiopeptidase A)EC 3.4.24.3 • Mr ca. 70 000 - 120 000 (collagenases) • CAS [9001-12-1]References and DefinitionsUnit definition: 1 U according to Wünsch (2) catalyzes the hydrolysis of 1 µmole 4-phenylazobenzyloxycarbonyl-L-prolyl-L-leucylglycyl-L-prolyl-D-arginine per minute at 25 °C, pH 7.1.References1. Bond, M.D. & van Wart, H.E. (1984) Biochemistry 23, 3077-30912. Wünsch, E. & Heidrich, H.G. (1963) Hoppe-Seyler's Z. Physiol. Chem. 333, 149-51
Collagenases from Clostridium histolyticum are proteolytic enzymes that cleave peptide bonds in the triple helical collagen molecules of human or animal tissue in situ. For this reason collagenases are widely used for isolation of various cell types by tissue dissociation.Collagenase NB 5 Sterile Grade is sterile according to European Pharmacopoeia. Collagenase NB 5 Sterile Grade is a crude collagenase that contains collagenolytic and additional enzymatic activities including clostripain and neutral protease. The balanced ratio of these activities ensures gentle and efficient tissue dissociation. SpecificationCollagenase activity: ≥0.10 U/mg (PZ acc. to Wünsch) Sterility: must comply Application Collagenase NB 5 Sterile Grade is suitable for dissociation of a broad variety of tissue types.Storage conditionsCollagenase NB 5 Sterile Grade is available as a lyophilized powder.It should be stored at +2 to +8℃ in a dry environment. Under these conditions the product is stable until the minimum shelf life stated on the certificate of analysis if repeated opening and closing of the vial is avoided. For storage of solutions please refer to “Stock solution”. Instructions for use: General Collagenase NB 5 Sterile Grade is suitable for isolation of a broad variety of cells from human or animal tissues. Tissue types include adipose tissue, cartilage, skin, placenta, and umbilical cord tissue. It can also be applied in cell culture for passaging, e.g. of embryonic stem cells. Tissue dissociationRecommended starting concentrations for selected applications: Adipose tissue (human, rodent): 0.2 – 0.3 PZ U/ml Cartilage (human, rodent): 0.3 – 0.4 PZ U/ml In general, the appropriate collagenase concentration depends on tissue type and origin as well as on the isolation procedure.Collagenase activity is at an optimum at 37℃ and pH 7.4.Stock solution Collagenase NB 5 Sterile Grade dissolves at a concentration of up to 150 mg/ml in all buffers which are commonly used for cell isolation. The enzyme solution must be constantly stored on ice. Since collagenase and some of the secondary proteases depend on calcium, it is recommended to use a buffer with ≥ 2 mM Ca²⁺. Absolutely no calcium chelating agents (e.g. EDTA) should be present at all. Reconstituted Collagenase NB 5 Sterile Grade can be aliquoted and stored at -20 °C. Aliquots are stable for 1 year if repeated freezing and thawing is avoided. Collagenase NB 5 Sterile Grade is sterile according to Ph. Eur. Therefore, 0.22 µm filtration is not necessary if sterile equipment and buffers are used. If 0.22 µm filtration is required, filters with low protein-binding properties (e.g. cellulose acetate, PVDF, or PES) are recommended.Working solutionTo prepare a working solution, the stock solution is diluted with buffer to achieve the required collagenase concentration. The working solution must be constantly stored on ice until use.Inactivation and inhibitors The dissociation process can be reduced, e.g. by cooling down or dilution of the enzyme solution.Collagenase is reversibly inactivated at high pH values and irreversibly inactivated at low pH values. Inhibitors of collagenase include cysteine or chelating agents like EDTA. Important note Collagenase NB 5 Sterile Grade is intended for research use only... Read More | Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 10.16 at 1.0 mg/ml for pure C3Molecular Weight187,000 Da (2 chains)General DescriptionRat C3 is purified from pooled normal rat serum. C3 is central to the activation of all three pathways of complement activation (Law, S.K.A. and Reid, KProtein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 10.16 at 1.0 mg/ml for pure C3Molecular Weight187,000 Da (2 chains)General DescriptionRat C3 is purified from pooled normal rat serum. C3 is central to the activation of all three pathways of complement activation (Law, S.K.A. and Reid, K.B.M. (1995)). Initiation of each pathway generates proteolytic enzyme complexes (C3 convertases) which are bound to the target surface. These enzymes cleave a peptide bond in C3 releasing the anaphylatoxin C3a and activating C3b. For a brief time (~60 µs) this nascent C3b is capable of reacting with and covalently coupling to hydroxyl groups on the target surface. Carbohydrates are the favored target, but protein hydroxyls and amino groups also react. This process of tagging the target surface with C3b is called opsonization. The reactive site in nascent C3b is a thioester (Tack B.J., et al. (1980); Pangburn M.K. and MüllerEberhard H.J. (1980)) and C3b is linked to the target through a covalent ester bond (an amide bond is formed if C3b is attached to amino groups). Most of the C3 activated during complement activation never attaches to the surface because its thioester reacts with water forming fluid phase C3b which is rapidly inactivated by factors H and I forming iC3b. Surface-bound C3b is necessary in all three pathways for efficient activation of C5 and formation of C5b-9 complexes that lyse the target cell membrane. Surface-bound C3b and its breakdown products iC3b and C3d are recognized by numerous receptors on lymphoid and phagocytic cells which use the C3b ligand to stimulate antigen presentation to cells of the adaptive immune system. The end result is an expansion of target-specific B-cell and T-cell populations.Physical Characteristics & StructureThe calculated molecular weight of rat C3 based on its amino acid sequence is 184,111daltons (without the signal peptide) and is similar to that of human C3 (185,000 daltons).The molecular weight of rat C3 as determined by SDS/polyacrylamide gel electrophoresis has been reported by Daha, M.R. et al., (1979) to be 187,000 daltons composed of two disulfide linked chains, alpha chain (123,000 daltons) and beta chain (76,000 daltons). The extinction coefficient of rat C3 (E1%/280nm = 10.16) is calculated based on its amino acid sequence using ProtParam and assumes all pairs of Cys residues form cystines (i.e. a pair of cysteine molecules are joined by a disulfide bond). The theoretical pI of rat C3 is 6.12. The normal plasma concentration of C3 inWistar rats has been reported to be 0.581mg/ml (Daha, M.R. et al., (1979)).FunctionThe biological functions of C3 are described above in the General Description section.GeneticsRat C3 chromosome location 9. The NCBI Gene ID number for rat C3 is 24232 and UniProt accession number is P01026.Precautions/Toxicity/HazardsThis protein is purified from animal plasma/serum and therefore precautions appropriate for handling any animal blood-derived product must be used.ReferencesLaw, S.K.A. and Reid, K.B.M. (1995) Complement 2nd Edition (ISBN 0199633568) Oxford University Press, Oxford.Tack BF, Harrison RA, Janatova J, Thomas ML, Prahl JW. (1980) Evidence for presence of an internal thiolester bond in third component of human complement. Proc Natl Acad Sci U S A. 77:5764-8.Pangburn M.K. and Müller-Eberhard H.J. (1980) Relation of putative thioester bond in C3 to activation of the alternative pathway and the binding of C3b to biological targets of complement. J Exp Med. 152:1102-14.Daha MR, Stuffers-Heiman M, Kijlstra A and Van ES LA. (1979) Isolation and characterization of the third component of rat complement. Immunology 36:63-70... Read More | Purity≥95% SDS-PAGE.FunctionStimulates growth of the cells in an autocrine manner. Mediates hormonal action on the growth of cancer cells | Purity> 95% by SDS-PAGE and HPLC analyses.FunctionGrowth factor that controls proliferation and cellular differentiation in the retina and bone formation. Plays a key role in regulating apoptosis during retinal development. Establishes dorsal-ventral positional information in the retina and Purity> 95% by SDS-PAGE and HPLC analyses.FunctionGrowth factor that controls proliferation and cellular differentiation in the retina and bone formation. Plays a key role in regulating apoptosis during retinal development. Establishes dorsal-ventral positional information in the retina and controls the formation of the retinotectal map (PubMed:23307924). Required for normal formation of bones and joints in the limbs, skull, digits and axial skeleton. Plays a key role in establishing boundaries between skeletal elements during development. Regulation of GDF6 expression seems to be a mechanism for evolving species-specific changes in skeletal strucutres. Seems to positively regulates differentiation of chondrogenic tissue through the growth factor receptors subunits BMPR1A, BMPR1B, BMPR2 and ACVR2A, leading to the activation of SMAD1-SMAD5-SMAD8 complex. The regulation of chondrogenic differentiation is inhibited by NOG (PubMed:26643732). Also involved in the induction of adipogenesis from mesenchymal stem cells. This mechanism acts through the growth factor receptors subunits BMPR1A, BMPR2 and ACVR2A and the activation of SMAD1-SMAD5-SMAD8 complex and MAPK14/p38... Read More | Purity≥95% SDS-PAGE.Endotoxin level<0.1 EU/µgFunctionMediates NK cell adhesion and triggers NK cell effector functions. Binds two different NK cell receptors: CD96 and CD226. These interactions accumulates at the cell-cell contact site, leading to the formation of a mature Purity≥95% SDS-PAGE.Endotoxin level<0.1 EU/µgFunctionMediates NK cell adhesion and triggers NK cell effector functions. Binds two different NK cell receptors: CD96 and CD226. These interactions accumulates at the cell-cell contact site, leading to the formation of a mature immunological synapse between NK cell and target cell. This may trigger adhesion and secretion of lytic granules and IFN-gamma and activate cytoxicity of activated NK cells. May also promote NK cell-target cell modular exchange, and PVR transfer to the NK cell. This transfer is more important in some tumor cells expressing a lot of PVR, and may trigger fratricide NK cell activation, providing tumors with a mechanism of immunoevasion. Plays a role in mediating tumor cell invasion and migration. Serves as a receptor for poliovirus attachment to target cells. May play a role in axonal transport of poliovirus, by targeting virion-PVR-containing endocytic vesicles to the microtubular network through interaction with DYNLT1. This interaction would drive the virus-containing vesicle to the axonal retrograde transport... Read More |