產(chǎn)品英文名稱：Porcine and Canine Insulin ELISA kit
產(chǎn)品中文名稱：豬/狗 胰島素ELISA檢測(cè)試劑盒
§ 檢測(cè)原理：ALPCO胰島素（豬/狗）ELISA是一個(gè)夾層式免疫檢測(cè)試劑盒。96孔微孔板涂有一層固相胰島素特異性鼠單克隆抗體。將標(biāo)準(zhǔn)物，對(duì)照品和樣品一并添加到微型板小孔中與山葵過(guò)氧化物酶標(biāo)記的單克隆抗體結(jié)合（配體物），胰島素分子被夾在固相和配體物之間。然后將微孔板放在搖床上室溫溫育2小時(shí)，700－900轉(zhuǎn)/分鐘。第一次溫育結(jié)束，用清洗緩沖液清洗小孔，去除未結(jié)合配體。添加TMB作用底物，放在微孔板搖床上第二次溫育15分鐘，700－900轉(zhuǎn)/分鐘。第二次溫育期間TMB和配體結(jié)結(jié)合物反應(yīng)呈藍(lán)色。第二次溫育結(jié)束，添加反應(yīng)終止液，調(diào)節(jié)分光光度計(jì)到450nm測(cè)定OD值。由于發(fā)光強(qiáng)度與樣品中胰島素含量成比例，因此可以根據(jù)OD值確定胰島素含量。
§ 檢測(cè)過(guò)程：各添加25μl標(biāo)準(zhǔn)物，對(duì)照物和樣品→加75μl配體物→微孔板搖床溫育2小時(shí)，700－900rpm→清洗緩沖液清洗6次→加100μl TMB作用物→微孔板搖床溫育15分鐘，700－900rpm→添加100μl反應(yīng)終止液→分光光度計(jì)450nm讀數(shù)
ALPCO Porcine and Canine Insulin ELISA kit英文簡(jiǎn)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.
