饥饿素(GHRL)检测试剂盒(酶联免疫吸附试验法)
ELISA Kit for Ghrelin (GHRL)
MTLRP; Growth Hormone-Releasing Peptide
- 编号CEA991Rb
- 物种Oryctolagus cuniculus (Rabbit,兔) 相同的名称,不同的物种。
- 实验方法竞争抑制
- 反应时长2h
- 检测范围123.5-10,000pg/mL
- 灵敏度最小可检测剂量小于等于52.3pg/mL.
- 样本类型Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- 下载 英文说明书 中文说明书
- 规格 48T96T 96T*5 96T*10 96T*100
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特异性
本试剂盒用于检测饥饿素(GHRL),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饥饿素(GHRL)(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) | 81-91 | 84 |
EDTA plasma(n=5) | 79-105 | 99 |
heparin plasma(n=5) | 85-92 | 88 |
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饥饿素(GHRL),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饥饿素(GHRL)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) | 89-98% | 92-101% | 78-103% | 78-90% |
EDTA plasma(n=5) | 85-101% | 89-97% | 97-105% | 88-98% |
heparin plasma(n=5) | 97-105% | 82-102% | 94-102% | 88-97% |
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。将饥饿素(GHRL)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:Oryctolagus cuniculus (Rabbit,兔) | 应用(仅供研究使用,不用于临床诊断!) |
CEA991Rb | 饥饿素(GHRL)检测试剂盒(酶联免疫吸附试验法) | Enzyme-linked immunosorbent assay for Antigen Detection. |
LMA991Rb | 饥饿素(GHRL)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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