| Description | FAD-dependent glucose dehydrogenase is an enzyme used as a regeneration cofactor to convert glucose and NAD(P) into NAD(P)H and gluconic acid.ORIGIN: Aspergillus oryzae CATALYSIS: SPECIFICATIONSActivity≥100 U/mg powderAppearanceYellow powder, lyophilizedStabilityStable for 36 months at -20 &#FAD-dependent glucose dehydrogenase is an enzyme used as a regeneration cofactor to convert glucose and NAD(P) into NAD(P)H and gluconic acid.ORIGIN: Aspergillus oryzae CATALYSIS: SPECIFICATIONSActivity≥100 U/mg powderAppearanceYellow powder, lyophilizedStabilityStable for 36 months at -20 ℃AdditivesNot AddedCHARACTERISTICSMolecular weight180,000 (Gel filtration)Isoelectric point6.5 Km (Glucose)15.0 × 10⁻³ M InhibitorsAg⁺, Hg²⁺ ActivatorTriton X-100 Opt. pH7.0Fig.1Opt. temperature55 ℃Fig.2Stable pH range4.0-7.0Fig.3Stable temp. rangebelow 40 ℃Fig.4Substrate specificity Table 1Mediator Preference Fig.5APPLICATIONS This enzyme is used for enzymatic determination of glucose in blood or urine by glucose sensor etc.Assay Method of Glucose Dehydrogenase (GDH-FAD)Principle1* phenadine methosulfate, 2* Nitorotetrazorium blueThe appearance of diformazan is measured at 570nm by spectrophotometry.Unit Definition One unit is defined as the enzyme quantity which produces 0.5 µmol of diformazan per minute under the conditions described below.ReagentsA. Triton X-100 Solution (5% Triton X-100 solution)Weigh 5.0 g of Triton X-100 and dissolve in 30 mL of deionized water with heating. After cooling, fill up to 100 mL with deionized water. (Expires after 1 month at room temperature)B. Working Solution (50 mmol/L PIPES-NaOH (pH 6.5) containing 0.5 % Triton X-100)Weigh 1.51 g of PIPES and dissolve in approx. 70mL of deionized water. Add 10 mL of Triton X-100 solution (A), then adjust the pH to 6.5 with 4N NaOH. Fill up to 100 mL with deionized water. (Expires after 1 month at 2~8℃)C. NTB Solution (6.6 mmol/L)Weigh 54 mg of Nitorotetrazorium blue and dissolve in 10 mL of deionized water. This solution should be kept in a light-proof tube to avoid exposure to light. (Expires after 14 days at 2~8℃)D. PMS Solution (3.0 mmol/L)Weigh 9 mg of phenazine methosulfate and dissolve in 10 mL of deionized water. This solution should be kept in a light-proof tube to avoid exposure to light. (Expires after 14 days at 2~8℃)E. Substrate Solution (1.0 mol/L Glucose)Weigh 3.6 g of D-glucose and dissolve in deionized water. Fill up to 20 mL with deionized water. (Expires after 14 days at room temperature)F. Enzyme DiluentWeigh 3.02 g of PIPES and 0.2 g of bovine serum albumin and 29.4 mg of CaCl₂·2H₂O in approx. 160 mL of deionized water. Add 4 mL of Triton X-100 solution (A), then adjust the pH to 6.5 with 4N NaOH. Fill up to 200 mL with deionized water. (Expires after 1 month at 2~8℃)G. Enzyme SolutionWeigh 25 mg of Glucose Dehydrogenase (GDH-FAD) and dissolve in chilled Enzyme Diluent (F). Enzyme Solution should be prepared so that the value of ΔOD/2minutes becomes in the range of 0.058±0.026.H. Mix SolutionMix 26 mL of Working Solution (B), 1 mL of NTB Solution (C), 2 mL of PMS Solution (D) and 1 mL of Substrate Solution (E). This solution should be kept in a light-proof tube to avoid exposure to light. (Expires 6 hours at 2~8℃)ProcedurePipette 3 mL of Mix Solution (H) into a disposable plastics cuvette (d=10 ~mm) and keep at 37±0.5 ℃ for 10 minutes. Then, pipette 0.1 mL of Enzyme Solution (E) into the cuvette and mix well immediately. Keep the reaction mixture at 37±0.5 ℃. Exactly at 3 minutes and 5 minutes after the addition of Enzyme Solution (E), measure the absorbances of the reaction mixture at 570 nm. (A3 and A5) As a blank, pipette Enzyme Diluent (F) into another disposable plastics cuvette (d=10 ~mm) instead of Enzyme Solution (E), and take the same procedure described above. (Ab3 and Ab5)Calculation2: Reaction time20.1: Half of millimolar extinction coefficient of diformazan at 570 nm3.1: Final volume of the reaction mixture0.1: Volume of Enzyme SolutionDm: Dilution multiple of Enzyme Solution... Read More | Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 0.631 at 1.0 mg/ml for pure C1qMolecular Weight400,000 Da (18 chains)General DescriptionRat C1q is purified from pooled normal rat serum. C1q is part of the C1 complex, which is the first complement component in the classical pathway of Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 0.631 at 1.0 mg/ml for pure C1qMolecular Weight400,000 Da (18 chains)General DescriptionRat C1q is purified from pooled normal rat serum. C1q is part of the C1 complex, which is the first complement component in the classical pathway of complement. The C1 complex is a non-covalent assembly of three different proteins (C1q, C1r, and C1s) bound together in a calcium-dependent complex. C1q has six extended arms with domains at the end of each arm that bind to the Fc domains of immunoglobulins such as IgG or IgM. When antibodies bind toantigens, forming immune complexes, they cluster allowing two or more of the six C1q arms to bind to the Fc domains of antibodies. Rat IgG2 is very efficient when compared to IgG1 in activating complement (Medgyesi, G.A et., al., 1981). This is in contrast to the human system in which IgG1 activates complement but not IgG2 (Redpath, S. et. al., 1998). The binding of multiple arms of C1q to immune complexes causes the two C1r proteins in the complex (protease zymogens) to auto-activate. The activated C1r proteases cleave and activate the two C1s protease zymogens in the complex. The activated C1s cleaves complement component C4 releasing C4a and initiating covalent attachment of C4b to the activating surface. Activated C1s also cleaves C2 and the larger fragment of C2 binds to the surface-attached C4b forming C4b,C2a, the C3/C5 convertase of the classical pathway.Rat IgG1 cannot activate complement whereas rat IgG2 does.Physical Characteristics & StructureThe apparent molecular weight of rat C1q as determined by gel filtration has been reported to be 400,000 by Veerhuis, R. et al., (1985) and is calculated to be 420,000 based on its amino acid sequence. Rat C1q is a high molecular weight complex of 18 polypeptide chains. Each of the six arms of rat C1q contains three chains, an A chain (~30,000 daltons), a B chain (~28,000 daltons) and a C chain (~26,000 daltons) as determined by SDS/polyacrylamide gel electrophoresis (Wing, M.G. et al., (1993)).FunctionThe biological functions of C1q are described above in the General Description and Physical Characteristics sections.ApplicationsRat C1q can be used to coat ELISA plates to capture and quantitate immune complexes in samples from rat models used for studying immune complex related diseases and conditions.GeneticsNCBI Gene ID numbers for rat C1q are: C1q A chain (298566), C1q B chain (29687), and C1q C chain (362634). The genes for C1q chains A, B and C are all located on chromosome 5. The UniprotKB primary accession numbers for rat C1q are: C1q A chain (P31720), C1q B chain (P31721), and C1q C chain (P31722).Precautions/Toxicity/HazardsThis protein is purified from animal plasma/serum and therefore precautions appropriate for handling any animal blood-derived product must be used.ReferencesMedgyesi, G.A et., Miklos, K., Kulics, J., Fust, G., and Gergely, J. Bazin, H. (1981). Classes and subclasses of rat antibodies: reaction with the antigen and interaction of the complex with the complement system. Immunology 43, 171-176.Redpath, S., Michaelsen, T., Sandlie, I. and Clark, M. R. (1998). Activation of complement by human IgG1 and human IgG3 antibodies against the human leucocyte antigen CD52. Immunology 93, 595–600.Veerhuis, R., Van Es, L.A. and Daha, M.R. (1985). In vivo degradation of rat C1q induced by intravenous injection of soluble IgG aggregates. Immunology 54, 801-810.Wing, M.G., Seilly, D. J., Bridgman, D.J. and Harrison, R.A. (1993). Rapid isolation and biochemical characterization of rat C1 and C1q. Molecular Immunology 30, 433-440... Read More | Inquire | Purity≥95% SDS-PAGE.Endotoxin level<0.1 EU/µgFunctionInhibits factor X (X(a)) directly and, in a Xa-dependent way, inhibits VIIa/tissue factor activity, presumably by forming a quaternary Xa/LACI/VIIa/TF complex. It possesses an antithrombotic action and also the ability to associate Purity≥95% SDS-PAGE.Endotoxin level<0.1 EU/µgFunctionInhibits factor X (X(a)) directly and, in a Xa-dependent way, inhibits VIIa/tissue factor activity, presumably by forming a quaternary Xa/LACI/VIIa/TF complex. It possesses an antithrombotic action and also the ability to associate with lipoproteins in plasma.Post-translationalO-glycosylated... Read More | Purity>95% SDS-PAGE.FunctionCytokine that binds to TNFRSF10A/TRAILR1, TNFRSF10B/TRAILR2, TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and possibly also to TNFRSF11B/OPG. Induces apoptosis. Its activity may be modulated by binding to the decoy receptors TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and TNFRSF11B/Purity>95% SDS-PAGE.FunctionCytokine that binds to TNFRSF10A/TRAILR1, TNFRSF10B/TRAILR2, TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and possibly also to TNFRSF11B/OPG. Induces apoptosis. Its activity may be modulated by binding to the decoy receptors TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and TNFRSF11B/OPG that cannot induce apoptosis... Read More |