胰岛素(INS)检测试剂盒(酶联免疫吸附试验法)

ELISA Kit for Insulin (INS)

  • 胰岛素(INS)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • 胰岛素(INS)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • 胰岛素(INS)检测试剂盒(酶联免疫吸附试验法) 实验结果图
  • CEA448Cp.jpg 标准曲线图
  • Certificate 通过ISO 9001、ISO 13485质量体系认证

特异性

本试剂盒用于检测胰岛素(INS),经检测与其它相似物质无明显交叉反应。
由于受到技术及样本来源的限制,不可能完成对所有相关或相似物质交叉反应检测,因此本试剂盒有可能与未经检测的其它物质有交叉反应。

回收率

分别于定值血清及血浆样本中加入一定量的胰岛素(INS)(加标样品),重复测定并计算其均值,回收率为测定值与理论值的比率。

样本 回收率范围(%) 平均回收率(%)
serum(n=5) 80-92 85
EDTA plasma(n=5) 88-102 93
heparin plasma(n=5) 87-98 92

精密度

精密度用样品测定值的变异系数CV表示。CV(%) = SD/mean×100
批内差:取同批次试剂盒对低、中、高值定值样本进行定量检测,每份样本连续测定20 次,分别计算不同浓度样本的平均值及SD值。
批间差:选取3个不同批次的试剂盒分别对低、中、高值定值样本进行定量测定,每个样本使用同一试剂盒重复测定8次,分别计算不同浓度样本的平均值及SD值。
批内差: CV<10%
批间差: CV<12%

线性

在定值血清及血浆样本内加入适量的胰岛素(INS),并倍比稀释成1:2,1:4,1:8,1:16的待测样本,线性范围即为稀释后样本中胰岛素(INS)含量的测定值与理论值的比率。

样本 1:2 1:4 1:8 1:16
serum(n=5) 97-105% 87-101% 85-94% 92-101%
EDTA plasma(n=5) 87-95% 84-99% 88-97% 80-93%
heparin plasma(n=5) 89-96% 91-104% 94-102% 94-101%

稳定性

经测定,试剂盒在有效期内按推荐温度保存,其活性降低率小于5%。
为减小外部因素对试剂盒破坏前后检测值的影响,实验室的环境条件需尽量保持一致,尤其是实验室内温度、湿度及温育条件。其次由同一实验员来进行操作可减少人为误差。

实验流程

1. 实验前标准品、试剂及样本的准备;
2. 加样(标准品及样本)50µL,
    加入50µL检测液A(临用前配制);
    37°C温育1小时。
3. 洗板3次;
4. 加检测溶液B100µL,37°C孵育30分钟;
5. 洗板5次;
6. 加TMB底物90µL,37°C孵育10-20分钟;
7. 加终止液50µL,立即450nm读数。

实验原理

本试剂盒应用竞争抑制酶联免疫分析法测定标本中待测物质水平。将胰岛素(INS)单克隆抗体包被微孔板,制成固相载体,往包被抗体的微孔中同时加入生物素标记的抗原和待测抗原(标准品或样本),待测抗原与生物素标记抗原对特异性抗体进行竞争结合。温育后经洗涤去掉未结合物,然后加入HRP标记的亲和素,经过温育和彻底洗涤后加入底物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最终的黄色。待测标本浓度越高,标记抗原和抗体的结合就越受到抑制,显色愈浅。显色的深浅与酶量呈正相关,而与样品中待测物质含量呈负相关。用酶标仪在450nm波长下测定吸光度(O.D.值),计算样品浓度。

相关产品

编号 适用物种:Capra hircus; Caprine (Goat,山羊) 应用(仅供研究使用,不用于临床诊断!)
CEA448Cp 胰岛素(INS)检测试剂盒(酶联免疫吸附试验法) Enzyme-linked immunosorbent assay for Antigen Detection.

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