產品英文名稱：Insulin ELISA Kit
產品中文名稱：人胰島素ELISA檢測試劑盒

人胰島素介紹：
胰島素是由胰島β細胞受內源性或外源性物質如核糖、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激素，由A、B鏈組成，共含51個氨基酸殘基。胰島素是機體內*降低血糖的激素，同時促進糖原、脂肪、蛋白質合成。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胰島素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，調節分光光度計到450nm測定OD值。由于發光強度與樣品中胰島素含量成比例，因此可以根據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

Insulin ELISA 英文簡述：
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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