| 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 | Inquire | Myelin Oligodendrocyte Glycoprotein Peptide (35-55), mouse, rat (MOG (35-55)) TFA is a minor component of CNS myelin. Myelin Oligodendrocyte Glycoprotein Peptide (35-55), mouse, rat TFA has encephalitogenic activity and induces T cell proliferative. Myelin Oligodendrocyte Glycoprotein Peptide Myelin Oligodendrocyte Glycoprotein Peptide (35-55), mouse, rat (MOG (35-55)) TFA is a minor component of CNS myelin. Myelin Oligodendrocyte Glycoprotein Peptide (35-55), mouse, rat TFA has encephalitogenic activity and induces T cell proliferative. Myelin Oligodendrocyte Glycoprotein Peptide (35-55), mouse, rat TFA induces Th1 cytokine response as well as relatively high levels of IgG antibodies. Myelin Oligodendrocyte Glycoprotein Peptide (35-55), mouse, rat TFA produces a relapsing-remitting neurological disease with extensive plaque-like demyelination... Read More | Inquire | Purity>97% by SDS-PAGE and HPLC analyses.FunctionMay be involved in macrophage-mediated cellular proliferation. It is mitogenic for fibroblasts and smooth muscle but not endothelial cells. It is able to bind EGF receptors with higher affinity than EGF itself and is a far more potent mitogen for Purity>97% by SDS-PAGE and HPLC analyses.FunctionMay be involved in macrophage-mediated cellular proliferation. It is mitogenic for fibroblasts and smooth muscle but not endothelial cells. It is able to bind EGF receptors with higher affinity than EGF itself and is a far more potent mitogen for smooth muscle cells than EGF. Also acts as a diphtheria toxin receptor.Background:Human HB-EGF (Heparin-Binding EGF-like growth factor) is a 12-16 kDa member of the EGF family of peptide growth factors (1-3). Also known as the DTR (diphtheria toxin receptor), it is further classified as a group 2 ErbB ligand based on its ability to activate both the EGF/ErbB1 and ErbB4 receptors (4, 5). HB-EGF is synthesized as a 208 amino acid (aa) type I transmembrane preproprecursor (1, 6). It contains a 19 aa signal sequence, a 43 aa prosegment, an 86 aa mature region (aa 63-148), an 11 aa juxtamembrane cleavage peptide, a 24 aa transmembrane segment, and a 25 aa cytoplasmic tail (aa 184-208). As an integral membrane protein, HB-EGF is expressed as a 19-27 kDa protein in mammalian cells (7-9). The variability in molecular weight (MW) is attributed to heterogeneity in glycosylation and/or the utilization of multiple proteolytic cleavage sites during maturation. Mature HB-EGF is a soluble peptide that arises from proteolytic processing of the transmembrane form. It possesses an EGF-like domain between aa 104-144, and a heparin-binding motif between aa 93‑113. Although the aa range for "mature" HB-EGF is typically stated to be Asp63-Leu148, potential N-terminal start (cleavage) sites also exist at Gly32, Arg73, Val74, Ser77 and Ala82 (8, 10-12). Thus, differential processing (in part) likely accounts for the 16-23 kDa range in MW noted for mammalian-derived mature HB-EGF. Proteases suggested to contribute to HB-EGF processing include TACE, MMP-3 and -7, ADAM-17 and ADAM-12 (11, 13-16). When expressed recombinantly in E.coli, HB-EGF (aa 73-148) runs at 14 kDa in SDS-PAGE; when expressed in Baculovirus, HB-EGF (aa 63-148, 77-148 and 32-148) runs at 18 kDa, 15 kDa, and 19 kDa respectively (8, 12, 17). Over aa 63-148, human HB-EGF- shares 76% and 73% aa sequence identity with rat and mouse HB-EGF, respectively (1, 18). Cells known to express HB-EGF include bronchial epithelium (19), visceral and vascular smooth muscle (20, 21), CD4+ T cells (22), cardiac muscle (23), glomerular podocytes (24), keratinocytes (13) and IL-10-secreting regulatory macrophages (25). As noted earlier, HB-EGF is known to bind to both 170 kDa EGFR and 180 kDa ErbB4, and through heterodimerization, ErbB2 (13, 26). Activity associated with ErbB4 binding appears to be limited to non-mitogenic actions, while EGFR binding induces both mitogenic and non-mitogenic activity... Read More |