产品英文名称：Insulin ELISA Kit
产品中文名称：人胰岛素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*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*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胰岛素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。*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

Insulin ELISA 英文简述：
The ALPCO Insulin ELISA kit is a FDA registered in vitro diagnostic tool for the quantification of human insulin in a clinical setting or research laboratory. Utilizing a dual-monoclonal antibody sandwich ELISA format, bi-level control set, and 96-well microplate comprising removable strips, a single Insulin ELISA kit has the performance characteristics and flexibility necessary to confidently measure up to 40 samples in duplicate. The shelf-life of the components and the resealable microplate pouch allow for convenient storage for future use if the entire kit is not needed at one time. Additional information regarding the Insulin ELISA kit can be accessed by clicking on the “Protocol" link or “Support" tab above.

PRINCIPLE OF THE ASSAY
The ALPCO Insulin ELISA is a sandwich type immunoassay. The 96-well microplate is coated with a monoclonal antibody specific for insulin. The standards, controls, and samples are added to the microplate wells with the Detection Antibody. The microplate is then incubated 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 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 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.
In the example below, a 5 pl curve fit was used to maximize the accuracy and precision of samples with low concentrations. However, the accuracy and precision of all models are limited at the lowest and highest ends of the detectable range due to the influence of individual laboratory conditions. As a result, caution should always be used when interpreting results where the analyte response becomes non-linear.

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