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人垂体中叶素(IMD)试剂盒2022年技术突破

阅读次数：1579次　发布时间：2022/11/16 16:59:39

人垂体中叶素（IMD）ELISA检测试剂盒

使用说明书

检测原理

试剂盒采用双抗体夹心法酶联免疫吸附试验（ELISA）。往预先包被人垂体中叶素（IMD）捕获抗体的包被微孔中，依次加入标本、标准品、HRP标记的检测抗体，经过温育并彻di洗涤。用底物TMB显色，TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成*终的黄色。颜色的深浅和样品中的人垂体中叶素（IMD）呈正相关。用酶标仪在450nm 波长下测定吸光度（OD 值），计算样品浓度。

Human Intermedin (IMD) ELISA Kit instruction

Intended use

This IMD ELISA kit is intended Laboratory for Research use only ａnd is not for use in diagnostic or therapeutic procedures. The Stop Solution changes the color from blue to yellow ａnd the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of IMD in the sample, this IMD ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples ａnd allow the operator to produce a standard curve of Optical Density versus IMD concentration. The concentration of IMD in the samples is then determined by comparing the O.D. of the samples to the standard curve.

洗板方法

1. 手工洗板：甩尽孔内液体，每孔加满洗涤液，静置1min后甩尽孔内液体，在吸水纸上拍干，如此洗板5次。

2. 自动洗板机：每孔注入洗液350μL，浸泡1min，洗板5次。

操作步骤

1. 从室温平衡20min后的铝箔袋中取出所需板条，剩余板条用自封袋密封放回4℃。

2. 设置标准品孔和样本孔，标准品孔各加不同浓度的标准品50μL；

3. 待测样本孔先加待测样本10μL，再加样本稀释液40μL；

4. 随后标准品孔和样本孔中每孔加入辣根过氧化物酶（HRP）标记的检测抗体100μL，用封板膜封住反应孔，37℃水浴锅或恒温箱温育60min。

5. 弃去液体，吸水纸上拍干，每孔加满洗涤液，静置1min，甩去洗涤液，吸水纸上拍干，如此重复洗板5次（也可用洗板机洗板）。

6. 每孔加入底物A、B各50μL，37℃避光孵育15min。

7. 每孔加入终止液50μL，15min内，在450nm波长处测定各孔的OD值。

Assay procedure

1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.

2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.

3. Add Sample: Add testing sample 10μl Then add sample diluent 40μl to testing sample well; Blank well doesn’t add anyting.

4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C.

5. Aspirate each well ａnd wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate ａnd blot it against clean paper towels.

6. Add chromogen solution A 50μl ａnd chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.

7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not

appear uniform, gently tap the plate to ensure thorough mixing.

8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.