| Description | ApplicationFor the detection of lactic acid content.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC 1.1.3.2Molecular weight: 42 kDa (SDS-PAGE)Isoelectric point: pH 4.6Km value: 7.5 × 10-4 M (L-Lactate)Inhibitors: Hg²⁺, Pb²⁺ Optimum pH: 6.0-7.0 ApplicationFor the detection of lactic acid content.Enzymatic propertiesSource: MicroorganismEnzymology Committee Number: EC 1.1.3.2Molecular weight: 42 kDa (SDS-PAGE)Isoelectric point: pH 4.6Km value: 7.5 × 10-4 M (L-Lactate)Inhibitors: Hg²⁺, Pb²⁺ Optimum pH: 6.0-7.0 Figure 1 Optimum temperature: 50℃ Figure 2pH stability: 6.0-8.5 (25℃,16h) Figure 3 Thermal stability: Stable below 50℃ (pH7.0, 30min) Figure 4Stability: -25 ~ -15℃ standing store for 12 monthsMore than 90% activity Figure 5 Enzyme activity definitionUnit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1µmol H2O2 per minute under the following conditions.Assay method for activity1. PrincipleThe amount of Quinoneimine dye produced by the reaction can be measured by spectrophotometer at 555 nm.2. Definition of enzyme activityUnit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1 µmol H2O2 per minute under the following conditions.3. Reagent preparationReagent I: 0.2 MpH 6.5 potassium phosphate buffer.Reagent II: 1kU/mL peroxidase (POD) solution.Reagent III: 50 mM4-AA solution.Reagent IV: 0.5 MDL- lactic acid solution, pH6.5.Reagent V: 50 mMTOOS solution.Enzyme diluent: 10 mMpH7.0 potassium phosphate buffer containing 10 µM FAD.Sample: Dilute the enzyme with enzyme diluent to 0.05-0.2U/mL.Prepare the reaction mixture as follows:Reagent I is 10 mlReagent II 0.25 mLReagent III 1.5 mLReagent IV is 5 mLReagent V 1.5 mLDouble steam water to 50 ml4. Operation procedure4.1 Add 1mL reaction mixture into 1mL colorimetric dish.4.2 Preheat the reaction mixture at 37 °C for 5min.4.3 Add 20µL of enzyme liquid to be measured and mix well.4.4 The reaction is measured at 37 °C at 555 nm and the absorbance change (∆As) within 1min is recorded.* Replace enzyme liquid with enzyme diluent, other steps are the same, the absorbance of the resulting solution is blank absorbance (∆Ab)∆A=∆As-∆Ab5. Vitality computing1.020: total volume of reaction liquid (mL);0.020: enzyme liquid volume (mL);1: optical path length (cm);1/2:1 mole of hydrogen peroxide to generate 1/2 mole of quinone imide dye;df: dilution ratio;C: Enzyme concentration (mg/mL);39.2: Under standard reaction conditions, the millimolar absorption coefficient of the color group at 555 nm (cm2/µmol)... Read More | 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 | Product Application：KNK437 has been used: as a heat shock factor 1 (HSF1) inhibitor to study its effects on the inhibition of viability and apoptosis activation in chemoresistant mice cells as an HSF1 inhibitor to study its effects on viability and apoptosis of colorectal cancer cells as a Product Application：KNK437 has been used: as a heat shock factor 1 (HSF1) inhibitor to study its effects on the inhibition of viability and apoptosis activation in chemoresistant mice cells as an HSF1 inhibitor to study its effects on viability and apoptosis of colorectal cancer cells as a heat shock protein 70 (HSP70) inhibitor to study its effects on glutamine-induced HSP70 and inflammatory mediator release... Read More | Store at +4°C. Store under desiccating conditions. The product can be stored for up to 12 months | Crystal phase: anatase/rutile ca. 80:20 |