產(chǎn)品說明 ：
產(chǎn)品貨號Reference: CAN-APN-5000
檢測方法Kit Type: Sandwich ELISA
規(guī)格Kit Size: 96-well break-apart microplate
靈敏度Sensitivity: 0.06 ng/mL
檢測樣本Sample Type: Human serum or plasma / 20 μL
檢測范圍Calibrator Range: 2–50 ng/mL
檢測時間Total Assay Time: 105 minutes
背景介紹：
脂聯(lián)素ELISA的原理是兩步夾心酶聯(lián)免疫分析。該方法使用了兩種高特異性的單克隆抗體：一種是將一種針對脂聯(lián)素的單克隆抗體固定在微板上，另一種是針對不同的脂聯(lián)素表位的單克隆抗體與生物素結(jié)合。在第一步中，樣品和標(biāo)準(zhǔn)品中的脂聯(lián)素結(jié)合到固定抗體和生物素化抗體上，從而形成一個三明治復(fù)合物。未結(jié)合的生物素化抗體通過洗滌去除。在第二步中，加入鏈霉親和素-HRP，它能特異性地與生物素化抗體結(jié)合。通過洗滌除去未結(jié)合的鏈霉親和素HRP。接下來，添加酶底物(TMB)。酶促反應(yīng)的顯色強度與脂聯(lián)素的濃度成正比。酶的反應(yīng)通過加入停止溶液而終止。在微孔板閱讀器上測量了450 nm處的吸光度。樣品和對照品中的脂聯(lián)素濃度可以從標(biāo)準(zhǔn)曲線的圖解中計算出來。
The principle of the adiponectin ELISA is a two-step sandwich enzyme immunoassay. The assay makes use of two highly specific monoclonal antibodies: A monoclonal antibody specific for adiponectin is immobilized onto the microplate and another monoclonal antibody specific for a different epitope of adiponectin is conjugated to biotin. During the first step, adiponectin present in the samples and standards is bound to the immobilized antibody and to the biotinylated antibody, thus forming a sandwich complex. Unbound biotinylated antibody is removed by a washing. In the second step, streptavidin-HRP is added, which binds specifically to bound biotinylated antibody. Unbound streptavidin-HRP is removed by washing. Next, the enzyme substrate (TMB) is added. The colour intensity of the enzymatic reaction is directly proportional to the concentration of adiponectin. The enzymatic reaction is terminated by the addition of stopping solution. The absorbance is measured on a microplate reader at 450 nm. The concentration of adiponectin in samples and controls can be calculated from of a plot of the standard curve, either graphically or by using immunoassay software.
