- ¥2828
- AATBIO
- 11304
- SigmaX-7375
- 2025年12月13日
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- 详细信息
- 文献和实验
- 技术资料
- 英文名：
荧光法黄嘌呤氧化酶检测试剂盒，红色荧光
- 供应商：
上海研卉生物科技有限公司
- 库存：
4W
- 规格：
200T
Xanthine oxidase (XO) is an enzyme that catalyzes the oxidation of hypoxanthine to xanthine and can further catalyze the oxidation of xanthine to uric acid. It plays an important role in the catabolism of purines. Xanthine oxidase is normally found in liver and jejunum. During severe liver damage, xanthine oxidase is released into blood, so a blood assay for XO is a way to determine if liver damage has happened. Xanthinuria is a rare genetic disorder where the lack of xanthine oxidase leads to high concentration of xanthine in blood and can cause health problems such as renal failure. The Amplite™ Fluorimetric Xanthine Oxidase Assay Kit provides a quick and ultrasensitive method for the measurement of xanthine oxidase activities. It can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. In the assay, xanthine oxidase catalyzes the oxidation of purine bases, hypoxanthine or xanthine to uric acid and superoxide , which spontaneously degrades to hydrogen peroxide (H2O2). The kit uses our Amplite™ Red substrate which enables a dual recordable mode. The fluorescent signal can be easily read by either a fluorescence microplate reader or an absorbance microplate reader. With the Amplite™ Xanthine Oxidase Assay Kit, we have detected as little as 0.15 mU/mL xanthine oxidase in a 100 µL reaction volume.
Platform
| Fluorescence microplate reader | |
| Excitation | 540nm |
| Emission | 590nm |
| Cutoff | 570nm |
| Recommended plate | Solid black |
Components
| Component A: Amplite™ Red (light sensitive) | 1 vial |
| Component B: Assay Buffer | 20 mL |
| Component C: Horseradish Peroxidase | 1 vial (lyophilized) |
| Component D: Xanthine | 100 µL (100 X) |
| Component E: Xanthine Oxidase Standard | 1 vial (200 mU, lyophilized) |
| Component F: DMSO | 1 vial (200 µL) |
Example protocol
At a glance
Protocol summary
- XO standards or test samples (50 µL)
- Add XO working solution (50 µL)
- Incubate at room temperature for 15 - 30 minutes
- Read fluorescence intensity at Ex/Em = 540/590 nm (cut off 570 nm)
Important notes
Thaw all the kit components to room temperature before starting the experiment.
Preparation of stock solution
1. Amplite™ Red stock solution (250X):
Add 40 µL of DMSO (Component F) into the vial of Amplite™ Red substrate (Component A). The stock solution should be used promptly. Note: The Amplite™ Red substrate is unstable in the presence of thiols such as dithiothreitol (DTT) and 2-mercaptoethanol. The final concentration of DTT or 2-mercaptoethanol in the reaction should be no higher than 10 µM. The Amplite™ Red substrate is also unstable at high pH (>8.5). Therefore, the reaction should be performed at pH = 7 – 8. The provided assay buffer, pH = 7.4, is recommended.
2. HRP stock solution (500X):
Add 100 µL of Assay Buffer (Component B) into the vial of Horseradish Peroxidase (Component C).
3. Xanthine Oxidase (XO) standard solution (1 U/mL)
Add 200 µL of Assay Buffer (Component B) into the vial of Xanthine Oxidase Standard (Component E).
Specifications
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文献和实验6-羟基嘌呤。与黄嘌呤一起，广泛存在于动植物体中，其核苷酸的肌苷酸是核酸的嘌呤核苷酸的前体。也是咖啡因的母体。次黄嘌呤可以通过腺嘌呤脱氨酶或亚硝酸的作用由腺嘌呤脱氨生成，也能通过核苷磷酸化酶使肌苷发生磷酸解而失成。通过黄嘌呤氧比酶可被氧化成尿酸，认为这是嘌呤碱的主要分解途经。
甲醛（去甲基作用的副产物）的形成。因此，易受洗涤剂、 巯基和一系列离子的干扰。与甲基化检测试剂盒类似，这些检测试剂盒对任何去甲基化酶都没有特异性，只能用纯化的蛋白进行检测。 组蛋白去甲基化酶检测试剂盒通过直接测定去甲基化产物的形成来规避这些问题，可确保： 组蛋白去甲基化酶检测试剂盒通过直接测定去甲基化产物的形成来规避这些问题，可确保： 灵敏度比基于甲醛的检测试剂盒更高（20 - 1,000 倍） 无硫醇、洗涤剂或离子干扰的更准确的数据 与核提取物或纯化蛋白的相容性（得益
达标记发出橙红色荧光，将两将荧光抗体按适当比例混合后，加在标本上（直接法）就分别形成抗原抗体复合物，发出黄绿色荧光的即抗A抗体结合部位，发出橙红色荧光的即抗B抗体结合的部位，这样就明确显示两种抗原的定位。 二、免疫酶法 免疫酶法是借助酶细胞化学等手段显示组织抗原（或抗体）的新技术，是在免疫荧光法的基础上发展起来的。已如前述，免疫荧光法具有操作简便、灵敏、特异性高、省时的优势，但同时也具有令人遗憾的不足，特别是荧光标本不能长期保存以及需要价格昂贵的荧光显微镜才能观察的问题
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