Technical indicators
Appearance: Colorless transparent liquid; Protein purity: ≥ 90%; Specific activity: ≥ 5 U/mL; β- N-acetylglucosamine glycosidase: ≤ 0.2%; α- Mannosidase: ≤ 0.05%; β- Galactosidase: ≤ 0.1%
Enzymatic properties
Source: Microorganisms; Classification: EC 3.2.1.51; Molecular weight: 55 kDa (SDS-PAGE); Isoelectric point: 6.2; Optimal pH: 7.5; Optimal temperature: 80 ℃; PH stability: 7.0-10.0 (25 ℃, 16 hours); Thermal stability: Stable below 90 ℃ (pH 8.0,30 min); Stability: Keep at -25~-15 ℃ for 12 months and maintain over 90% activity.
purpose
Used for calibration of reagents such as fucosidase, research and development of quality control products, and large-scale preparation.
Definition of enzyme activity
Unit enzyme activity is defined as hydrolyzing 1% per minute under the following conditions μ The amount of enzyme required for mol CNP-AFU.
Reagent preparation
Reagent I: 20mM pH 8.0 Tris HCl.
Reagent II: 10mM CNP-AFU solution (dissolved in DMSO).
Enzyme diluent: 20mM pH 8.0 Tris HCl.
Operating Steps
1. Add 940 to a 1mL colorimetric dish μ Reagents I and 50 for L μ L Reagent II.
2. Preheat the reaction mixture at 37 ℃ for 5 minutes.
3. Add 10 μ L enzyme solution to be tested, mix well.
4. Measure the reaction at 37 ℃ at 405nm and record the absorbance change within 1 minute（ Δ As).
*Replace the enzyme solution with enzyme diluent, and the other steps are the same. The absorbance of the obtained solution is blank absorbance（ Δ Ab) Δ A= Δ As- Δ Ab
