Ⅳ型胶原(COL4)检测试剂盒(酶联免疫吸附试验法)

ELISA Kit for Collagen Type IV (COL4)

Type-IV Collagen

  • Ⅳ型胶原(COL4)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • Ⅳ型胶原(COL4)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • Ⅳ型胶原(COL4)检测试剂盒(酶联免疫吸附试验法) 实验结果图
  • SEA180Mu.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Ⅳ型胶原(COL4)，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Ⅳ型胶原(COL4)（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) 88-102 96
EDTA plasma(n=5) 86-102 93
heparin plasma(n=5) 80-89 81

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Ⅳ型胶原(COL4)，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Ⅳ型胶原(COL4)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) 88-104% 85-99% 80-93% 79-91%
EDTA plasma(n=5) 82-96% 80-99% 83-101% 97-105%
heparin plasma(n=5) 78-98% 88-96% 86-95% 93-101%

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）100µL，37°C孵育1小时；
3. 吸弃，加检测溶液A100µL，37°C孵育1小时；
4. 洗板3次；
5. 加检测溶液B100µL，37°C孵育30分钟；
6. 洗板5次；
7. 加TMB底物90µL，37°C孵育10-20分钟；
8. 加终止液50µL，立即450nm读数。

实验原理

将Ⅳ型胶原(COL4)抗体包被于96孔微孔板中，制成固相载体，向微孔中分别加入标准品或标本，其中的Ⅳ型胶原(COL4)与连接于固相载体上的抗体结合，然后加入生物素化的Ⅳ型胶原(COL4)抗体，将未结合的生物素化抗体洗净后，加入HRP标记的亲和素，再次彻底洗涤后加入TMB底物显色。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。颜色的深浅和样品中的Ⅳ型胶原(COL4)呈正相关。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：Mus musculus (Mouse，小鼠) 应用(仅供研究使用，不用于临床诊断！)
SEA180Mu Ⅳ型胶原(COL4)检测试剂盒(酶联免疫吸附试验法) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA180Mu Ⅳ型胶原(COL4)等多因子检测试剂盒(流式荧光发光法) FLIA Kit for Antigen Detection.

参考文献

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