| Description | ApplicationIt is used in the research and development of enzymatic glycosylated albumin reagents and mass formulation.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC1.5.3Molecular weight: 55kDa (SDS-PAGE)Isoelectric point: 6Km value: 5.0×10 ⁻⁴ M (Fructosyl-ApplicationIt is used in the research and development of enzymatic glycosylated albumin reagents and mass formulation.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC1.5.3Molecular weight: 55kDa (SDS-PAGE)Isoelectric point: 6Km value: 5.0×10 ⁻⁴ M (Fructosyl-Ala)Inhibitors: Hg ²⁺, Pb ²⁺ Optimal pH: 7.7 Figure 1Optimum temperature: 42℃ Figure 2pH stability: 5.0-9.5 (25℃, 16h) Figure 3Thermal stability: Stable below 40℃ (pH8.0, 30min) Figure 4Stability: -25 ~ -15℃ standing store for 12 monthsMore than 90% activity Figure 5Protective agent: glycerin, trehaloseAssay method for activity1. Principle 2. Definition of enzyme activityUnit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1µmol H2O2 per minute under the following conditions.3. Reagent preparationReagent I: 1M potassium phosphate buffer, pH8.0Reagent II: 1kU/mL PODReagent III: 50mM TOOS solutionReagent IV: 50mM 4-AA solutionReagent V: 200mM Glycated alanineEnzyme diluent: 20mM Tris-HCl, pH8.0Prepare the reaction mixture as follows:Reagent I: 10mLReagent II: 0.1mLReagent III: 1mLReagent IV: 1mLReagent V: 10mLDouble steam water set volume to 100mL4. Operation procedure4.1 Add 980µL reaction mixture into 1mL colorimetric dish.4.2 Incubate at 37°C for 5 minutes.4.3 Add 20 µL of enzyme solution to be tested to the reaction mixture.4.4 Reaction at 37°C, the absorbance change (∆As) of the sample within 1min is detected by spectrophotometer at 555nm.* Blank control measurement method: Use 20 µL of enzyme dilution solution instead of the enzyme solution to be tested, and measure the absorbance change (ΔAb) of the sample within 1 minute.∆A=∆As-∆Ab5. Vitality computingVt: Total volume of reaction liquid (1.0 mL);Vs: Enzyme liquid volume (0.02 mL);t: Reaction time (1 min);df: Dilution ratio;C: Enzyme concentration (mg/mL);1.0: Optical path length (cm);1/2: 1 mole hydrogen peroxide to generate 1/2 mole quinone imide dye;39.2: Under standard reaction conditions, the millimolar absorption coefficient of the color group at 555nm (cm² /µmol)... Read More | DescriptionApolipoprotein E (ApoE) is present in the brain and is mainly produced by astrocytes. It is a 299 amino acid glycoprotein of 34kDa. It is present in all classes of lipoproteins except LDL (low-density lipoprotein). APOE gene has three alleles, such as APOE ε3, APOE ε4and APOE DescriptionApolipoprotein E (ApoE) is present in the brain and is mainly produced by astrocytes. It is a 299 amino acid glycoprotein of 34kDa. It is present in all classes of lipoproteins except LDL (low-density lipoprotein). APOE gene has three alleles, such as APOE ε3, APOE ε4and APOE ε2. It is located on human chromosome 19q13.Preparation instructionsFormLyophillized from a 0.2 µm filtered solution in 20 mM sodium phosphate, pH 7.8.Principle... 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 | Aprotinin is a competitive serine protease inhibitor that inhibits trypsin,chymotrypsin,kallikrein and plasmin.Aprotinin forms stable complexes with and blocks the active sites of enzymes. Binding is reversible with most aprotinin,protease complexes and dissociating at pH >10 or <3. Effective Aprotinin is a competitive serine protease inhibitor that inhibits trypsin,chymotrypsin,kallikrein and plasmin.Aprotinin forms stable complexes with and blocks the active sites of enzymes. Binding is reversible with most aprotinin,protease complexes and dissociating at pH >10 or <3. Effective concentration is equimolar with protease.Recombinant aprotinin is expressed in E. Coli, and purified with HPLC. It contains no animal-derived components. This is a recombinant form of bovine lung aprotinin, which is traditionally isolated from bovine lung by methods involving fractional precipitation, gel filtration, and ion exchange chromatography. UNIT DEFINITION:A conversion factor for Aprotinin is: 1 EPU = 1 USP Aprotinin Unit = 1800 KIU... Read More | Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description: 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hingePurity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description: 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hinge region with a hydrophobic cleft that is exposed upon Ca2+ binding. S100B is a 91 amino acid (aa) protein, after removal of the initial methionine, and is found as homodimers of 10.4 kDa monomers. Human S100B shares 99%, 98%, 100%, 99% and 97% aa sequence identity with mouse, rat, rabbit, equine and bovine S100B, respectively. Within the S100 family, human S100B shows the highest aa identity (59%) with S100A1. S100B is expressed primarily by astrocytes and oligodendrocytes in the central nervous system, and by Schwann cells in the peripheral nervous system. Ca2+-bound S100B interacts in vitro with at least 20 cytoplasmic proteins, including several structural molecules such as tubulin and GFAP. It can inhibit the phosphorylation of these kinase substrates and others such as tau and neuromodulin. Astrocytes can secrete S100B, which then acts in a cytokine-like manner. Nanomolar concentrations of S100B are secreted constitutively, promote proliferation, and are neurotrophic and anti-apoptotic. Blood levels of S100B reflect extracellular concentrations within the nervous system, and are elevated in Down’s syndrome, Alzheimer’s disease and Tourette’s syndrome, metabolic stress, acute brain injury and brain tumors. Micromolar concentrations of S100B can be destructive and pro-apoptotic; they induce the expression of iNOS, COX-2, IL-1, IL‑6 and TNF-alpha by microglia, astrocytes or neurons. Most extracellular actions of S100B can be mediated by RAGE (receptor for advanced glycation end products), which is also a receptor for other S100 proteins... Read More |