产品英文名称：Porcine and Canine Insulin ELISA kit

产品中文名称：猪/狗 胰岛素ELISA检测试剂盒

§ 检测原理：ALPCO胰岛素（猪/狗）ELISA是一个夹层式免疫检测试剂盒。96孔微孔板涂有一层固相胰岛素特异性鼠单克隆抗体。将标准物，对照品和样品一并添加到微型板小孔中与山葵过氧化物酶标记的单克隆抗体结合（配体物），胰岛素分子被夹在固相和配体物之间。然后将微孔板放在摇床上室温温育2小时，700－900转/分钟。第一次温育结束，用清洗缓冲液清洗小孔，去除未结合配体。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。第二次温育期间TMB和配体结结合物反应呈蓝色。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25μl标准物，对照物和样品→加75µl配体物→微孔板摇床温育2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

ALPCO Porcine and Canine Insulin ELISA kit英文简述：

The ALPCO Porcine and Canine Insulin ELISA kit is for the quantification of porcine or canine insulin concentrations in serum and plasma samples. Utilizing a dual-monoclonal antibody sandwich format, bi-level control set, and 96-well microplate comprising removable strips, a single Porcine and Canine 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 Porcine and Canine Insulin ELISA kit can be accessed by clicking on the “Protocol” link or “Support” tab above.

INTENDED USE

The ALPCO Insulin (Porcine/Canine) ELISA is for the quantitative determination of insulin in Porcine/ Canine serum and plasma.

PRINCIPLE OF THE ASSAY

The ALPCO Insulin (Porcine/Canine) ELISA is a sandwich type immunoassay. Mouse monoclonal antibodies specific for insulin are immobilized to the 96-well microplate as the solid phase. Standards, controls, and samples are added to the appropriate wells with a horseradish peroxidase enzyme labeled monoclonal antibody (Conjugate), resulting in insulin molecules being sandwiched between the solid phase and the Conjugate. After incubation on a microplate shaker at room temperature, the microplate wells are washed with Wash Buffer to remove unbound Conjugate. TMB Substrate is added to each well, and the microplate is again incubated on a microplate shaker at room temperature. During the second incubation, a blue color results from TMB Substrate reacting with bound Conjugate in the wells. Stop Solution is added and this stops the reaction and changes the color from blue to yellow. The optical density (OD) is measured by microplate reader at 450 nm with a reference wavelength of 620 nm. The intensity of the color generated is directly proportional to the amount of insulin in the sample.