线粒体开放阅读框12S rRNA-C(MOTS-c)检测试剂盒(酶联免疫吸附试验法,小样本)

Mini Samples ELISA Kit for Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c)

特异性

本试剂盒用于检测线粒体开放阅读框12S rRNA-C(MOTS-c)检测试剂盒(酶联免疫吸附试验法,小样本),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线粒体开放阅读框12S rRNA-C(MOTS-c)检测试剂盒(酶联免疫吸附试验法,小样本)(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)94-10197
EDTA plasma(n=5)97-105101
heparin plasma(n=5)83-9592

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线粒体开放阅读框12S rRNA-C(MOTS-c)检测试剂盒(酶联免疫吸附试验法,小样本),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线粒体开放阅读框12S rRNA-C(MOTS-c)检测试剂盒(酶联免疫吸附试验法,小样本)含量的测定值与理论值的比率。

样本1:21:41:81:16
serum(n=5)82-96%91-99%79-92%91-102%
EDTA plasma(n=5)80-97%97-105%78-94%79-104%
heparin plasma(n=5)93-101%82-96%96-105%99-105%

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)25µL后,立即加入检测溶液A25µL,
    37°C孵育1小时;
3. 甩干,洗板3次;
4. 加检测溶液B50µL,37°C孵育30分钟;
5. 洗板5次;
6. 加TMB底物50µL,37°C孵育10-20分钟;
7. 加终止液25µL,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。将线粒体开放阅读框12S rRNA-C(MOTS-c)检测试剂盒(酶联免疫吸附试验法,小样本)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。

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参考文献

杂志参考文献
Pediatric Diabetes Circulating MOTS‐c levels are decreased in obese male children and adolescents and associated with insulin resistance[Pubmed:29691953]
Rapid Commun Mass SpectromDevelopment of a mass spectrometry‐based detection method for the mitochondrion‐derived peptide MOTS‐c in plasma samples for doping control purposes[Pubmed: 30394592]
DIABETOLOGIAAdiponectin treatment improves insulin resistance in mice by regulating the expression of the mitochondrial-derived peptide MOTS-c and its response to exercise via …[Pubmed: 32880686]
Induction of hyperandrogenism and insulin resistance differentially modulates ferroptosis in uterine and placental tissues of pregnant rats[]
FASEB JOURNALNeural cell©\derived plasma exosome protein abnormalities implicate mitochondrial impairment in first episodes of psychosis[33454965]
Biochim Biophys Acta Mol Basis DisMOTS-c interacts synergistically with exercise intervention to regulate PGC-1¦Á expression, attenuate insulin resistance and enhance glucose metabolism in mice via?¡­[33722744]
researchsquareEvolution of Mitochondrial-Derived Peptides, Humanin and MOTSc and Changes in Insulin Sensitivity During Early Gestation[]
Mol PsychiatryAbnormal levels of mitochondrial proteins in plasma neuronal extracellular vesicles in major depressive disorder[34471251]
Pharmacol ResThe Mitochondrial-Derived Peptide MOTS-c Relieves Hyperglycemia and Insulin Resistance in Gestational Diabetes Mellitus[34798268]
Ann NeurolSARS‐CoV‐2 and mitochondrial proteins in neural‐derived exosomes of COVID‐19[Pubmed:35285072]
J Clin MedEvolution of Mitochondrially Derived Peptides Humanin and MOTSc, and Changes in Insulin Sensitivity during Early Gestation in Women with and without Gestational …[Pubmed:35683389]