| Description | ApplicationOxidized glycosylated amino acids, used in the development and mass preparation of enzymatic glycosylated hemoglobin reagents.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC1.5.3Molecular weight: 60 kDa (SDS-PAGE)Isoelectric point: 6.4Km value: 4.0×10-3M (ApplicationOxidized glycosylated amino acids, used in the development and mass preparation of enzymatic glycosylated hemoglobin reagents.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC1.5.3Molecular weight: 60 kDa (SDS-PAGE)Isoelectric point: 6.4Km value: 4.0×10-3M (fructosyl-Val-His)Inhibitors: Hg²⁺, Pb²⁺ Optimum pH: 6.5-7.5 Figure 1Optimum temperature: 37℃ Figure 2pH stability: pH 6.5-9.5 (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, trehalose Assay method for activity1. Principle The resulting Quinoneiminedye can be detected at 555nm.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 conditions3. Reagent preparationReagent I: 0.1M potassium phosphate buffer, pH8.0Reagent II: 1kU/mLPOD.Reagent III: 50mMTOOS solution (1.477gTOOS dissolved in 100mLUP water).Reagent IV: 50 mm4-AA solution (1.016g4-AA dissolved in 100mLUP water).Reagent V: 200mM glycosylated valine.Sample: Diluted with enzyme diluent 20mMTris-HCl, pH8.0.Prepare the reaction mixture as follows:Reagent I: 10mLReagent II: 0.1mLReagent III: 1mLReagent IV: 1mLReagent V: 10mLDouble steam water set volume to 100mLSample: Diluted with enzyme diluent 20mMTris-HCl, pH8.0.4. Operation procedure4.1 Add 980µL reaction mixture to 1mL colorimetric dish.4.2 Incubate at 37°C for 5min.4.3 Add 20µL of enzyme solution to the reaction mixture.4.4 Reaction at 4.37°C, the absorbance change (∆As) within 1min of the sample is detected by spectrophotometer at 555nm.* Replace the enzyme solution to be tested with enzyme diluent and determine the absorbance change (∆Ab) of the sample within 1min.∆A=∆As-∆Ab5. Vitality computing Vt: total volume of reaction liquid (1.0mL);Vs: enzyme liquid volume (0.02mL);t: Reaction time (1min);df: dilution ratio;C: Enzyme concentration (mg/mL);1.0: optical path length (cm);1/2:1 mole of hydrogen peroxide to generate 1/2 mole of quinone imide dye;39.2: Under standard reaction conditions, the millimolar absorption coefficient of the color group at 555nm (cm2/µmol)... Read More | Inquire | Product DescriptionEndo F1 cleaves Asparagine-linked high mannose and some hybrid oligosaccharides. Core fucosylation reduces the activity by 50 fold. Endoglycosidase F1 will hydrolyze sulfate containing high-mannose chains. It cleaves between the two N-acetylglucosamine residues in the Product DescriptionEndo F1 cleaves Asparagine-linked high mannose and some hybrid oligosaccharides. Core fucosylation reduces the activity by 50 fold. Endoglycosidase F1 will hydrolyze sulfate containing high-mannose chains. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact.Molecular weight 32,000 daltonsContents60 µl aliquot of enzyme (1 U) in 20 mM Tris-HCl, pH 7.5Included with 20 µL and 60 µL pack sizes:5x Reaction Buffer – 250 mM sodium phosphate, pH 5.5Specific ActivityDefined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured Ribonuclease B (RNase B) in 1 minute at 37°C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved RNase B migrates faster).FormulationThe enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, pH 7.5StabilitySeveral days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly. SpecificityEndo F1 cleaves Asparagine-linked high mannose or hybrid oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Quality & PurityEndo F1 is tested for contaminating protease as follows: 10 µg of denatured BSA is incubated at 37°C for 24 hours with 2 µl of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. The production host strain has been extensively tested and does not produce any detectable glycosidases.Directions for use1. Add up to 200 µg of glycoprotein to an Eppendorf tube. Adjust to 38 µl final volume with de-ionized water.2. Add 10 µl 5x Reaction Buffer 5.53. Add 2.0 µl of Endo F1 to the reaction. Incubate 1 hour or more at 37°C.Monitor cleavage by SDS-PAGE... Read More | Purity:>98%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：Heme oxygenase (HMOX) is the rate limiting enzyme in heme catabolism. It cleaves heme to biliverdin, carbon monoxide, and iron. The biliverdin is subsequently converted to bilirubin by biliverdin reductase. Purity:>98%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：Heme oxygenase (HMOX) is the rate limiting enzyme in heme catabolism. It cleaves heme to biliverdin, carbon monoxide, and iron. The biliverdin is subsequently converted to bilirubin by biliverdin reductase. The mechanism of HMOX is unique in that heme serves as the substrate of the enzyme and as the prosthetic group for the activation of iron-bound O2. HMOX activity is highest in spleen where senescent erythrocytes are sequestered and destroyed. Two isoforms, HMOX1 and HMOX2, are expressed in most tissues. HMOX1 is an inducible enzyme in response to heme, heavy metals, oxidative stress, cytokines, and many drugs. Whereas HMOX2 displays a constitutive expression. HMOX1 is expressed mainly in spleen, liver, and kidney, and HMOX2 is prominently expressed in the brain and testes. The increased expression of HMOX1 levels is related to a variety of pathological states, where it functions as a cytoprotective molecule through its by products. HMOX1 also plays important roles in the regulation of cell proliferation, differentiation, and apoptosis... Read More | Tyrosine decarboxylase catalyzes the removal of the carboxyl group from tyrosine to produce tyramine and carbon dioxide. Pyridoxal 5'-phosphate is a necessary cofactor. By using the apoenzyme prepared from cells grown on a vitamin B6 deficient medium pyridoxal phosphate may be determined. The Tyrosine decarboxylase catalyzes the removal of the carboxyl group from tyrosine to produce tyramine and carbon dioxide. Pyridoxal 5'-phosphate is a necessary cofactor. By using the apoenzyme prepared from cells grown on a vitamin B6 deficient medium pyridoxal phosphate may be determined. The HOLOenzyme may be used to determine tyrosine, phenylalanine and dihydroxyphenylalanine either manometrically or colorimetrically.L-Tyrosine decarboxylase apoenzyme from Streptococcus faecalis has been used in a study to purify and characterize tyrosine decarboxylase and aromatic-L-amino-acid decarboxylase.L-Tyrosine decarboxylase apoenzyme from Streptococcus faecalis has also been used in a study to investigate the stereospecificity of sodium borohydride reduction of tyrosine decarboxylase... Read More |