牛肺病(ECP)说明书定性.doc

牛肺病（ECP）酶联免疫ELISA） 试剂盒使用说明书本试剂仅供研究使用实验原理本试剂盒用双抗夹心法测定中。用纯化的包被微孔板，制成固相抗，，经过彻底洗涤后底物TMB显色。TMB在酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。用酶标仪在nm波长下测定吸光度（OD值）-20℃保存，但应避免反复冻融操作步骤μl。然后在待测样品孔μl，然后再加待测样品10μl。加样将样品加于酶标板孔底部，尽量不触及孔壁，轻轻混匀，分钟甩干μl，空白孔除外。 μl，再加入显色剂B 50μl，轻轻震荡混匀，37℃避光显色15分钟 终止：每孔加终止液50μl，终止反应。450nm波长依序测量各孔的度（OD值）≥1.00; 阴性对照平均值≤0.10 临界值（CUT OFF）计算：临界值=阴性对照孔平均值+0.15 阴性判定：样品OD值 临界值（CUT OFF）者为肺病（ECP）阴性 阳性判定：样品OD值≥ 临界值（CUT OFF）者为肺病（ECP）阳性 注意事项 1．操作严格按照说明书进行，本试剂不同批号组分不得混用。 2．试剂盒从冷藏环境中取出应在室温平衡15-30分钟后方可使用，酶标包被板开封后如未用完，板条应装入密封袋中保存。 3．浓洗涤液会有析出，稀释时可在水浴中加温助溶1．；2-8。 Bovine ECP FOR RESEARCH USE ONLY Drug Names Generic Name：ELISA Kit. Purpose This kit allows for the determination of ECP concentrations in Bovine serum, and other biological fluids. Principle of the assay The kit assay ECP level in the sample，use Purified antibody to coat microtiter plate wells, make solid-phase antibody, then add ECP to wells, Combined With ECP, after washing and removing non-combinative antidody and other components ,then Combined ECP antibody which with HRP labeled become antibody – antigen - enzyme- antibody complex, after washing Completely, Add TMB substrate solution,, TMB substrate becomes blue color At HRP enzyme-catalyzed, reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. Compared with the CUTOFF value, according to this to judge ECP exist in the sample or not. Materials provided with the kit Materials provided with the kit 48determinations 96 determinations Storage User manual 1 1 Closure plate membrane 2 2 Sealed bags 1 1 Microelisa stripplate 1 1 2-8℃ Negative control 0.5ml×1 bottle 0.5ml×1 bottle 2-8℃ Positive control 0.5ml×1 bottle 0.5ml×1 bottle 2-8℃ HRP-Conjugate reagent 3ml×1 bottle 6ml×1 bottle 2-8℃ Sample diluent 3ml×1 bottle 6ml×1 bottle 2-8℃ Chromogen Solution A 3ml×1 bottle 6ml×1 bottle 2-8℃ Chromogen Solution B 3ml×1 bottle 6ml×1 bottl