中英文介绍丨艾美捷丙二醛(MDA)elisa检测试剂盒使用

Principle of the Assay：

This kit was based on Competitive-ELISA detection method. The microtiter plate provided in this kit has been pre-coated with target. During the reaction, target in the sample or standard competes with a fixed amount of target on the solid phase supporter for sites on the Biotinylated Detection Antibody specific to target. Excess conjugate ａnd unbound sample or standard are wash ed from the plate, ａnd HRP-Streptavidin (SABC) is added to each microplate well ａnd incubated. Then TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution ａnd the color change is measured spectrophotometrically at a wavelength of 450nm. The concentration of target in the samples is then determined by comparing the OD of the samples to the standard curve.

Precautions

1. To inspect the validity of experiment operation ａnd the appropriateness of sample dilution proportion, pilot experiment using standards ａnd a small number of samples is recommended.

2. After opening ａnd before using, keep plate dry.

3. Before using the kit, spin tubes ａnd bring down all components to the bottom of tubes.

4. Storage TMB reagents avoid light.

5. Washing process is very important, not fully wash easily cause a false positive ａnd high background.

6. Duplicate well assay is recommended for both standard ａnd sample testing.

7. Don’t let microplate dry at the assay, for dry plate will inactivate active components on plate.

8. Don’t reuse tips ａnd tubes to avoid cross contamination.

9. Please do not mix the reagents in different kits of our company. Do not mix reagents from other manufacturers.

10. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary.

艾美捷丙二醛(MDA)elisa检测试剂盒#EKF60157检测原理：

该试剂盒基于竞争ELISA检测方法。该试剂盒中提供的微量滴定板已预先涂有目标在反应过程中，样品或标准品中的靶标与固相载体上固定量的靶标竞争用于靶特异性的生物素化检测抗体上的位点。洗涤过量的缀合物和未结合的样品或标准品并将HRP链亲和素（SABC）加入每个微孔板孔中并孵育。然后TMB基板溶液添加到每个井中。通过加入硫酸溶液和颜色变化终止酶底物反应在450nm的波长下用分光光度法测量。然后通过以下方式确定样品中目标的浓度将样品的OD与标准曲线进行比较。

丙二醛(MDA)elisa检测试剂盒操作步骤：

步骤1：在预涂板上分别设置标准、试样、对照（空白）孔，然后记录它们的位置。

第2步：在每个孔中加入50ul标准品或样品。立即向每个孔中加入50ul生物素标记的抗体，轻轻敲击

平板以确保充分混合，然后在37°C下孵育45分钟。

清洗步骤：抽吸并清洗盘子3次。

步骤3：向每个孔中加入100ul SABC工作溶液，并在37°C下孵育30分钟。

清洗步骤：抽吸并清洗盘子5次。

第4步：添加90ul TMB基质溶液。在37°C下培养10-20分钟。

步骤5：添加50ul停止溶液。在450nm下立即读取并计算

注意事项：

1.为了检验实验操作的有效性和样品稀释比例的适当性，建议使用标准品和少量样品进行中试。

2.打开后和使用，请保持盘子干燥。

3.在使用试剂盒之，旋转试管，并将所有组件降到试管底部。

4.TMB试剂应避光保存。

5.洗涤过程非常重要，不完全洗涤容易造成假阳性和高背景。

6.对于标准测试和样品测试，建议使用双孔分析法。

7.在测定时不要让微孔板干燥，因为干燥的平板会使平板上的活性成分失活。

8.不要重复使用尖端和管道，以避免交叉污染。

9.请不要将试剂混合在我们公司的不同试剂盒中。不要混合其他制造商的试剂。

10.为了确保准确的结果，在培养步骤中需要适当地粘附平板密封剂。

来源：https://www.amyjet.com/products/EKF60157.shtml