肌酸脒基水解酶（CR）

肌酸脒基水解酶

Creatine amidinohydrolase | CR

rp216175-5KU

5KU

BioReagent

≥90%(SDS-PAGE)

分子量： 48 kDa 酶学委员会编号： EC 3.5.3.3 物理外观：Lyophilized powder 水解肌酸，用于酶法肌酐试剂的研发和大量配制。 酶学性质（注意+、2+为上标） 来源： 微生物 酶学委员会编号： EC 3.5.3.3 分子量： 48 kDa (SDS-PAGE) 等电点： 4.6 Km值： 8.81×10-3M(Creatine) 抑制剂： Ag ⁺ ，Cu²⁺，Hg²⁺ 最适pH： 8.0 图1 最适温度： 45℃ 图2 pH稳定性： 4.5-10.0(25°C,16h) 图3 热稳定性： 50°C以下稳定(pH7.5,30min) 图4 保护剂： EDTA及糖类物质 活性测定方法 1、原理 2、酶活定义 单位酶活定义为在下述条件下，每分钟催化反应生成1 μmol肌酸完全转化为肌氨酸所需的酶量。 3、试剂准备 试剂I：0.1M肌酸溶液。 试剂II：将2.0g DAB溶解在100mL二甲亚砜中，并加入15mL浓盐酸。 酶稀释液：50mMPBS，pH7.5，样品用酶稀释液将酶稀释至2.0-3.0U/mL。 4、操作步骤 1.取1mL试剂I于5mL EP管中，37°C预热5min。 2.加入0.1mL酶液，混匀。 3.37°C水浴反应10min后，加入2.0mL试剂II终止反应，并在25°C放置20min。 4.使用紫外分光光度计，测定反应液在435nm处吸光度值。 5.酶稀释液代替酶液作为空白对照。 5、 活力计算 3.1：反应液总体积（mL）； 0.1：酶液体积（mL）； 10：反应时间（min）； 1.0：光路长度(cm)； df：稀释倍数； C：酶浓度(mg/mL)； 0.321：黄色染料在435nm处毫摩尔吸光系数(cm²/μmol)。 Molecular weight: 48 kDa Enzymology Committee Number: EC 3.5.3.3 Physical appearance: Lyophilized powder Hydrolyzed creatine, used in the development and mass formulation of enzymatic creatinine reagents. Enzymatic properties (note that + and 2+ are superscripts) Source: Microorganism Enzymology Committee No. : EC 3.5.3.3 Molecular weight: 48 kDa (SDS-PAGE) Isoelectric point: 4.6 Km value: 8.81×10-3M(Creatine) Inhibitors: Ag ⁺ , Cu²⁺, Hg²⁺ Optimal pH: 8.0 图1 Optimum temperature: 45℃ 图2 pH stability: 4.5-10.0(25°C,16h) 图 3 Thermal stability: Stable below 50°C (pH7.5,30min) 图 4 Protective agents: EDTA and sugar Assay method for activity 1. Principle Urea (Urea) and DAB (rho - dimethylaminobenzaldenhyde) reaction to form the yellow dye amount shall be tested by spectrophotometer at 435 nm. 2. Definition of enzyme activity Unit enzyme activity is defined as the amount of enzyme required to catalyze a reaction to produce 1 μmol of creatine per minute for complete conversion to sarcosine under the following conditions. 3. Reagent preparation Reagent I: 0.1M creatine solution. Reagent II: 2.0gDAB was dissolved in 100mL dimethyl sulfoxide and 15mL concentrated hydrochloric acid was added. Enzyme diluent: 50mMPBS, pH7.5, enzyme diluent was used to dilute the sample to 2.0-3.0U/mL. 4. Operation procedure 1. Take 1mL reagent I and preheat it in 5mLEP tube at 37°C for 5min. 2. Add 0.1mL of enzyme liquid and mix well. 3. After 10min reaction in a water bath at 37°C, 2.0mL reagent II was added to terminate the reaction and placed at 25°C for 20min. 4. Use an ultraviolet spectrophotometer to determine the absorbance value of the reaction liquid at 435nm. 5. Enzyme diluent was used as blank control instead of enzyme liquid. 5. Vitality computing 3.1: total volume of reaction liquid (mL); 0.1: enzyme liquid volume (mL); 10: Reaction time (min); 1.0: optical path length (cm); df: dilution ratio; C: Enzyme concentration (mg/mL); 0.321: millimolar absorption coefficient of yellow dye at 435nm (cm²/μmol).