Insulin Ultrasensitive ELISA Kit
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产品英文名称:Insulin Ultrasensitive ELISA Kit
产品中文名称:人胰岛素超灵敏ELISA检测试剂盒
人胰岛素介绍:
胰岛素是由胰岛β细胞受内源性或外源性物质如核糖、乳糖、葡萄糖、胰 高血糖素、精氨酸等的刺激由胰腺朗格汉斯小岛所分泌的一种蛋白质激素,由A、B链组成,共含51个氨基酸残基。胰岛素是机体内唯一降低血糖的激素,同时促进糖原、脂肪、蛋白质合成。外源性胰岛素主要用来进行糖尿病治疗,糖尿病患者早期使用胰岛素和超强抗氧化剂如(口服虾青素、注射用硫辛酸等)有望出现较长时间的蜜月期,胰岛素注射不会有成瘾和依赖性。
§ 检测原理:ALPCO人胰岛素超灵敏ELISA是一个夹层式免疫检测试剂盒。96孔微孔板涂有一层胰岛素特异性单克隆抗体。将标准物,对照品和样品一并添加到微型板小孔中与检测抗体混合。然后将微孔板放在摇床上温育1小时,700-900转/分钟。第一次温育结束,用清洗缓冲液清洗小孔并烘干。添加TMB作用底物,放在微孔板摇床上第二次温育15分钟,700-900转/分钟。第二次温育结束,添加反应终止液,调节分光光度计到450nm测定OD值。由于发光强度与样品中胰岛素含量成比例,因此可以根据OD值确定胰岛素含量。
§ 检测过程:各添加25 μl标准物,对照物和样品→加100µl检测抗体→微孔板摇床温育1小时,700-900rpm→清洗缓冲液清洗6次→加100µl TMB作用物→微孔板摇床温育15分钟,700-900rpm→添加100µl反应终止液→分光光度计450nm读数
Ultrasensitive Insulin ELISA英文简述:
INTENDED USE
The ALPCO Ultrasensitive Insulin ELISA is designed for the quantitative determination of insulin
in human serum and plasma.
PRINCIPLE OF THE ASSAY
The ALPCO Ultrasensitive Insulin ELISA is a sandwich type immunoassay. The 96-well microplate is coated with a monoclonal antibody specific for insulin. The standards, samples, and the control are added to the microplate wells with the Detection Antibody. The microplate is then incubated at room temperature on a microplate shaker at 700-900 rpm. After the first incubation is complete, the wells are washed with Wash Buffer and blotted dry. TMB Substrate is added, and the microplate is incubated a second time at room temperature on a microplate shaker at 700-900 rpm. Once the second incubation is complete, Stop Solution is added, and the optical density (OD) is measured by a spectrophotometer at 450 nm. The intensity of the color generated is directly proportional to the amount of insulin in the sample.
CALCULATION OF RESULTS
Construct a standard curve from the standards. The Zero Standard should be used as a blank with its average value subtracted from each well. It is recommended to use a software program to calculate the standard curve and to determine the concentration of the samples.
The ALPCO Ultrasensitive Insulin ELISA is a ligand binding assay, with responses exhibiting a sigmoidal relationship to the analyte concentration. Currently accepted reference models for such curves use a 4 or 5 parameter logistic (pl) fit, as these models optimize the accuracy and precision across a greater range. Although cubic spline and other models are acceptable methods, they generally show less intra-assay accuracy and precision at the low and high ends of the range.
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