Product category
產(chǎn)品英文名稱:Insulin Ultrasensitive ELISA Kit
產(chǎn)品中文名稱:人胰島素超靈敏ELISA檢測試劑盒
人胰島素介紹:
胰島素是由胰島β細胞受內(nèi)源性或外源性物質(zhì)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漢斯小島所分泌的一種蛋白質(zhì)激素,由A、B鏈組成,共含51個氨基酸殘基。胰島素是機體內(nèi)*降低血糖的激素,同時促 進糖原、脂肪、蛋白質(zhì)合成。外源性胰島素主要用來進行糖尿病治療,糖尿病患者早期使用胰島素和*抗氧化劑如(口服蝦青素、注射用硫辛酸等)有望出現(xiàn)較長 時間的蜜月期,胰島素注射不會有成癮和依賴性。
§ 檢測原理:ALPCO人胰島素超靈敏ELISA是一個夾層式免疫檢測試劑盒。96孔微孔板涂有一層胰島素特 異性單克隆抗體。將標準物,對照品和樣品一并添加到微型板小孔中與檢測抗體混合。然后將微孔板放在搖床上溫育1小時,700-900轉/分鐘。*次溫育 結束,用清洗緩沖液清洗小孔并烘干。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/分鐘。第二次溫育結束,添加反應終止 液,調(diào)節(jié)分光光度計到450nm測定OD值。由于發(fā)光強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。
§ 檢測過程:各添加25 μl標準物,對照物和樣品→加100µl檢測抗體→微孔板搖床溫育1小時,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反應終止液→分光光度計450nm讀數(shù)
Ultrasensitive Insulin ELISA英文簡述:
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.