Application Oxidized glycosylated amino acids, used in the development and mass preparation of enzymatic glycosylated hemoglobin reagents. Enzymatic properties Source: Microorganism Enzymology Committee Number: EC1.5.3 Molecular weight: 60 kDa (SDS-PAGE) Isoelectric point: 6.4 Km value: 4.0×10-3M (fructosyl-Val-His) Inhibitors:
Hg²⁺, Pb²⁺ Optimum pH: 6.5-7.5 Figure 1 Optimum temperature: 37℃ Figure 2 pH stability: pH 6.5-9.5 (25℃,16h) Figure 3 Thermal stability: Stable below 40℃ (pH8.0, 30min) Figure 4 Stability: -25 ~ -15℃ standing store for 12 months More than 90% activity Figure 5 Protective agent: glycerin, trehalose
Assay method for activity 1. Principle
The resulting Quinoneiminedye can be detected at 555nm. 2. Definition of enzyme activity Unit 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 preparation Reagent I: 0.1M potassium phosphate buffer, pH8.0 Reagent 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: 10mL Reagent II: 0.1mL Reagent III: 1mL Reagent IV: 1mL Reagent V: 10mL Double steam water set volume to 100mL Sample: Diluted with enzyme diluent 20mMTris-HCl, pH8.0. 4. Operation procedure 4.1 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-∆Ab 5. 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).
规格
产品名称
果糖基肽氧化酶（FPOX）
别名
果糖基肽氧化酶
英文别名
Fructosyl-peptide oxidase; FPOX
规格或纯度
BioReagent, EnzymoPure™, ≥90%(SDS-PAGE), ≥ 4 U/mg
CAS编号和信息
rp216180
分子类型
酶
产品线
生物产品
储存与运输
物理形态
冻干(Lyophilized)
浓度
≥ 4 U/mg
复溶
50–100 mM Tris-HCl缓冲液， pH 7.5–8.5
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输
稳定性与储存
Store at -20℃ long term (12 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.
ZJ25F0521340
危险品化学品经营许可证（带存储）