饥饿素(GHRL)检测试剂盒(酶联免疫吸附试验法,小样本)

Mini Samples ELISA Kit for Ghrelin (GHRL)

MTLRP; Growth Hormone-Releasing Peptide

  • 饥饿素(GHRL)检测试剂盒(酶联免疫吸附试验法,小样本) 产品包装(模拟)
  • 饥饿素(GHRL)检测试剂盒(酶联免疫吸附试验法,小样本) 产品包装(模拟)
  • 饥饿素(GHRL)检测试剂盒(酶联免疫吸附试验法,小样本) 实验结果图
  • MEA991Mu.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) 90-101 94
EDTA plasma(n=5) 88-95 91
heparin plasma(n=5) 90-98 95

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) 79-96% 83-101% 83-90% 93-101%
EDTA plasma(n=5) 91-105% 80-93% 87-99% 93-101%
heparin plasma(n=5) 78-92% 98-105% 80-97% 87-94%

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。

试剂盒内容

试剂名称 数量 试剂名称 数量
96孔板(预包被) 1 96孔板覆膜 4
标准品 2 标准品稀释液 1×20mL
检测溶液A 1×60µL 检测稀释液A 1×6mL
检测溶液B 1×60µL 检测稀释液B 1×6mL
TMB底物 1×4.5mL 终止液 1×3mL
浓洗涤液 (30 × concentrate) 1×10mL 说明书 1

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)25µL,37°C温育1小时;
3. 吸弃,加检测溶液A25µL,37°C孵育1小时;
4. 洗板3次;
5. 加检测溶液B25µL,37°C孵育30分钟;
6. 洗板5次;
7. 加TMB底物25µL,37°C孵育10-20分钟;
8. 加终止液20µL,立即450nm读数。

Mini Samples ELISA Kit for Ghrelin (GHRL)

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。

增值服务

相关产品

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参考文献

杂志 参考文献
Archives of Oral Biology Expression of ghrelin in human salivary glands and its levels in saliva and serum in Chinese obese children and adolescents [PubMed: 21075359]
Department of Medical Biochemistry and Clinical Biochemistry Presence of adropin, nesfatin-1, apelin-12, ghrelins and salusins peptides in the milk, cheese whey and plasma of dairy cows [ScienceDirect: S0196978113000533]
Journal of Physiology and Pharmacology Postnatal administration of 2-oxoglutaric acid improves articular and growth plate cartilages and bone tissue morphology in pigs prenatally treated with dexamethasone [PubMed: 23211309]
Gene The modifying effects of fish oil on fasting ghrelin mRNA expression in weaned rats [PubMed: 22842192]
Hormone Research in Paediatrics Changes in Ghrelin and Obestatin Levels before and after a Meal in Children with Simple Obesity and Anorexia [Pubmed: 23774845]
Surgical Innovation Gastric supply manipulation to modulate ghrelin production and enhance vascularization to the cardia: proof of the concept in a porcine model [Pubmed:25294792]
Obesity Surgery Embolization of Arterial Gastric Supply in Obesity (EMBARGO): an Endovascular Approach in the Management of Morbid Obesity. Proof of the Concept in the Porcine Model [Pubmed:25511751]
Obes Surg Effect of Sleeve Gastrectomy Plus Side-to-Side Jejunoileal Anastomosis for Type 2 Diabetes Control in an Obese Rat Model [PubMed: 26202420]
Surg Innov Gastric Supply Manipulation to Modulate Ghrelin Production and Enhance Vascularization to the Cardia Proof of the Concept in a Porcine Model [PubMed: 25294792]
Nutrients.  Elevation of Fasting Ghrelin in Healthy Human Subjects Consuming a High-Salt Diet: A Novel Mechanism of Obesity? [Pubmed:27240398]
World Journal of Gastroenterology Effects of sleeve gastrectomy with jejuno-jejunal or jejuno-ileal loop onglycolipid metabolism in diabetic rats. [pubmed:27621579]
Ginekologia Polska Increased serum ghrelin in preeclampsia: is ghrelin a friend or a foe? [pubmed:27321099]
Gynecological Endocrinology The association between serum ghrelin levels and large-for-gestational-age fetuses in patients with gestational diabetes mellitus [pubmed:27791436]
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