热休克蛋白70(HSP70)检测试剂盒(酶联免疫吸附试验法)

ELISA Kit for Heat Shock Protein 70 (HSP70)

DnaK; 70 Kilodalton Heat Shock Proteins

  • 热休克蛋白70(HSP70)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • 热休克蛋白70(HSP70)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • 热休克蛋白70(HSP70)检测试剂盒(酶联免疫吸附试验法) 实验结果图
  • SEA873Mi.jpg 标准曲线图
  • Certificate 通过ISO 9001、ISO 13485质量体系认证

特异性

本试剂盒对检测热休克蛋白70(HSP70)具有较高的灵敏度。
经检测牛、山羊、绵羊之间有100%的交叉反映。

回收率

分别于定值血清及血浆样本中加入一定量的热休克蛋白70(HSP70)(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) 82-103 91
EDTA plasma(n=5) 80-101 85
heparin plasma(n=5) 91-105 98

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热休克蛋白70(HSP70),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热休克蛋白70(HSP70)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) 96-105% 78-95% 89-96% 79-101%
EDTA plasma(n=5) 90-104% 95-102% 94-103% 78-103%
heparin plasma(n=5) 86-94% 78-98% 89-103% 82-99%

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

将热休克蛋白70(HSP70)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,其中的热休克蛋白70(HSP70)与连接于固相载体上的抗体结合,然后加入生物素化的热休克蛋白70(HSP70)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热休克蛋白70(HSP70)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:Bos taurus; Bovine (Cattle,牛), Capra hircus; Caprine (Goat,山羊), Ovis aries; Ovine (Sheep,绵羊) 应用(仅供研究使用,不用于临床诊断!)
SEA873Mi 热休克蛋白70(HSP70)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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