计算溶液所需的质量、体积或浓度。
黄嘌呤氧化酶活性检测试剂盒（紫外比色法）
有货
为什么选择此级别
BioReagent,用于显微镜,适用于分析 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
黄嘌呤氧化酶 (Xanthione oxidase, XOD) 是一种专一性不高，既能催化次黄嘌呤生成黄嘌呤，进而生成尿酸，又能直接催化黄嘌呤生成尿酸的酶，是一种含钼、非血红素铁、无机硫化物、FAD 的黄素酶，通常存在于哺乳动物的心，肺，肝脏等组织中。虽然血中 XO 活性通常很低，但肝损伤会导致XO 释放到血液中。对肝损害的诊断具有特异性的意义。原理：XOD 催化黄嘌呤产生尿酸，尿酸在 290nm 下有特征吸收峰。
产品组分
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使用方法
1.样品的准备：
(1) 细菌、细胞样品的制备：细菌或培养细胞：先收集细菌或细胞到离心管内，离心后弃上清；按照细菌或细胞数量：提取液体积(mL)为 500：1 的比例(建议 500万细菌或细胞加入 1 mL 提取液)，适当吹打，以充分裂解细胞；8000 g 4℃离心 10 min，取上清，置冰上待测。
(2) 组织样品的制备：组织：按照组织质量(g)：提取液 体积(mL)为 1：5~10 的比例(建议称取约 0.1 g 组织，加入 1 mL 提取液 )，进行冰浴匀浆。8000 g 4℃离心 10min，取上清，置冰上待测。
(3) 血清（浆）样品：直接检测。
2. 测定步骤：
(1) 分光光度计预热 30 min 以上，调节波长至 295 nm。
(2) 按照每10 mL缓冲液，加入60 μL底物的比例配XOD 工作液，现用现配。
(3) 测定前将 XOD 工作液在 37℃(哺乳动物)或 25℃(其它物种)水浴 20 min 以上。
(4) 在 1 mL 石英比色皿中加入下列试剂:
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3. XOD 活性计算公式
(1) 血清(浆)XOD 计算：
单位的定义：每毫升血清(浆)每分钟催化产生 1 µmol 尿酸定义为一个酶活力单位。
XOD (U/mL) = [ΔA×V 反总 ÷(ε×d)×10⁶]÷V 样 ÷T = 2.424×ΔA(2) 组织、细菌或细胞中XOD 计算：
按样本蛋白浓度计算,单位的定义：每 mg 组织蛋白每分钟催化产生 1 µmol 尿酸定义为一个酶活力单位。
OD (U/mg prot) = [ΔA×V 反总 ÷(ε×d)×10⁶] ÷( V 样 ×C pr )÷T = 2.424×ΔA÷C pr 按样本鲜重计算,单位的定义：每g 组织每分钟催化产生 1µmol 尿酸定义为一个酶活力单位。
XOD (U/g 鲜重) = [ΔA×V 反总 ÷(ε×d)×10⁶] ÷(W×V 样 ÷V 样总 )÷T = 2.424×ΔA÷W 按细菌或细胞密度计算,单位的定义：每一万个细菌或细胞每分钟催化产生1 µmol 尿酸定义为一个酶活力单位。
XOD (U/104 cell) = ΔA×V 反总 ÷(ε×d)×10⁶] ÷(500× V 样 ÷V 样总 )÷T = 4.848×10⁻³×ΔA 注： V 反总 ：反应体系总体积，1.035× 10⁻³ L；ε：尿酸摩尔消光系数，0.122×10⁵ L/mol/cm；d：比色皿光径，1 cm； V 样 ：加入样本体积，0.035 mL； V 样总 ：加入提取液体积，1 mL；T：反应时间，1 min；W：样本质量，g； C pr ：样本蛋白质浓度，mg/mL；500：细胞或细菌总数，500 万； 10⁶ ：单位换算系数，1 mol= 10⁶ µmol。
Xanthine oxidase (XOD) is an enzyme with relatively low substrate specificity. It catalyzes hypoxanthine to form xanthine and subsequently uric acid; it can also directly catalyze xanthine to produce uric acid. As a flavoenzyme containing molybdenum, non‑heme iron, inorganic sulfide and FAD, XOD is commonly found in mammalian tissues such as heart, lung and liver. Although XO activity in blood is normally low, liver injury causes XO to be released into the bloodstream, which is of specific significance for the diagnosis of hepatic damage.
Principle: XOD catalyzes xanthine to generate uric acid, which exhibits a characteristic absorption peak at 290 nm.
Product Components
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Operating Procedures
1. Sample Preparation
(1) Preparation of bacterial or cell samples
Collect bacteria or cultured cells into centrifuge tubes, centrifuge and discard the supernatant. Add extraction solution at a ratio of 500 : 1 (number of bacteria or cells : volume of extraction solution in mL; recommended: add 1 mL extraction solution to 5 million bacteria or cells). Pipette up and down adequately for complete cell lysis. Centrifuge at 8000 g for 10 min at 4 °C. Transfer the supernatant and keep it on ice for subsequent assay.
(2) Preparation of tissue samples
Add extraction solution to tissue at a mass-volume ratio of 1 g tissue : 5-10 mL extraction solution (recommended: weigh approximately 0.1 g tissue and add 1 mL extraction solution). Perform ice-bath homogenization. Centrifuge at 8000 g for 10 min at 4 °C. Transfer the supernatant and keep it on ice for subsequent assay.
(3) Serum / plasma samples: measure directly without pretreatment.
2. Assay Protocol
(1) Warm-up the spectrophotometer for no less than 30 min and set the wavelength to 295 nm.
(2) Prepare XOD working solution freshly before use: add 60 μL substrate per 10 mL assay buffer.
(3) Incubate the XOD working solution in a water bath for ≥20 min before measurement: 37 °C for mammalian samples or 25 °C for samples from other species.
(4) Pipette reagents into a 1 mL quartz cuvette as shown below:
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Mix thoroughly and start timing immediately. Zero the instrument using the blank tube. Record the initial absorbance A₁ at 295 nm and absorbance A₂ after 1 min. Calculate ΔA = A-A₁.
3. Calculation of XOD Activity
(1) Calculation for serum / plasma XOD
Unit definition: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the production of 1 µmol uric acid per minute per milliliter of serum/plasma.
XOD (U/mL) = [ΔA × Vtotal reaction ÷ (ε × d) × 10⁶] ÷ Vsample ÷ T = 2.424 × ΔA
(2) Calculation for XOD in tissues, bacteria or cells
Calculation based on sample protein concentration
Unit definition: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the production of 1 µmol uric acid per minute per milligram of tissue protein.
XOD (U/mg prot) = [ΔA×Vtotal reaction ÷ε×d) × 10⁶] ÷ (Vsample × Cpr) ÷ T = 2.424 × ΔA ÷ Cpr
Calculation based on sample fresh weight
Unit definition: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the production of 1 µmol uric acid per minute per gram of tissue fresh weight.
XOD (U/g fresh weight) = [ΔA × Vtotal reaction ÷ (ε × d) × 10⁶] ÷ (W × Vsample ÷ Vtotal sample) ÷ T = 2.424 × ΔA ÷ W
Calculation based on sample fresh weight
Unit definition: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the production of 1 µmol uric acid per minute per gram of tissue fresh weight.
XOD (U/g fresh weight) = [ΔA × Vtotal reaction ÷ (ε × d) × 10⁶] ÷ (W × Vsample ÷ Vtotal sample) ÷ T = 2.424 × ΔA ÷ W
Calculation based on bacterial or cell count
Unit definition: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the production of 1 µmol uric acid per minute per 10 000 bacteria or cells.
XOD (U/10⁴ cell) = [ΔA × Vtotal reaction ÷ (ε × d) × 10⁶] ÷ (500 × Vsample ÷ Vtotal sample) ÷ T = 4.848 × 10⁻³× ΔA
Notes:
Vtotal reaction: total volume of reaction system, 1.035×1010⁻³ L
ε: molar extinction coefficient of uric acid, 1.22×10⁴ L·mol⁻¹·cm⁻¹
d: optical path length of cuvette, 1 cm
Vsample : volume of added sample, 0.035 mL
Vtotal sample: total volume of added extraction solution, 1 mL
T: reaction time, 1 min
W: sample weight (g)
Cpr: protein concentration of sample (mg/mL)
500: total number of bacteria/cells, 5 million
10⁶: unit conversion factor (1 mol = 10⁶ µmol)
规格
规格或纯度
BioReagent, 用于显微镜, 适用于分析
英文名称
Xanthine Oxidase Activity Assay Kit (UV Colorimetric Method)
应用
细胞代谢, 酶活性检测
检测方法
比色法
定量&半定量
定量
检测仪器
紫外-可见光分光光度计
检测波长
A295
敏感性
Light-sensitive
检测时长
30 min
样本体积
20μL
储存与运输
储存条件
2-8°C储存
运输条件
低温运输
稳定性与储存
各组分在相应的储存条件下保质期1年。
组分与储存
组分货号 | 组分名称 | 外观 | 50 T | 储存 | 单次测试使用量 |
X1522837A | 提取液 | 液体 | 60 mL | 2-8℃. | 按需 |
X1522837B | 缓冲液 | 液体 | 60 mL | 2-8℃. | 1 mL |
X1522837C | 底物 | 液体 | 360 μL | 2-8℃. 避光保存 | 6 μL |
技术文档
高级数据
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F1039953 |
分析证书 | X1522837 |
技术文档和文章
溶液计算器
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