人神经调节蛋白1nrg-1酶联免疫分析.docVIP

人神经调节蛋白1(NRG-1)酶联免疫盒使用说明书实验原理本试剂盒应用双抗体夹心法测定中水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入，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。颜色的深浅和样品中的呈正相关。用酶标仪在nm波长下测定吸光度（OD值），计算样品浓度。 试剂盒组成 标准酶板稀释液稀释液-20℃保存，但应避免反复冻融操作步骤 5号标准品 150μl的原倍标准品加入150μl标准品稀释液 6pg/mL 4号标准品 150μl的5号标准品加入150μl标准品稀释液 3pg/mL 3号标准品 150μl的4号标准品加入150μl标准品稀释液 1.5pg/mL 2号标准品 150μl的3号标准品加入150μl标准品稀释液 0.75pg/mL 1号标准品 150μl的2号标准品加入150μl标准品稀释液 加样：分别设空白孔（空白对照孔不加样品及酶标试剂，其余各步操作相同）、标准孔、待测样品孔μl，待测样品孔中先加样品稀释液40μl，然后再加待测样品10μl（样品最终稀释度为5倍）。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B50μl，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）计算以标准物的浓度为横坐标，OD值为纵坐标，在坐标纸上绘出标准曲线，根据样品的OD值由标准曲线查出相应的浓度；再乘以；或用标准物的浓度与OD值计算出标准曲线的直线回归方程式，将样品的OD值代入方程式，计算出样品浓度，再乘以，即为样品的实际浓度。 浓洗涤液会有析出，稀释时可在水浴中加温助溶乘以1．；2-8。 NRG-1 FOR RESEARCH USE ONLY Assay range：0.3pg/mL - 12pg/mL 96 determinations Purpose For the quantitative in vitro determination of NRG-1 concentrations in Human serum, cell culture supernates and other biological fluids Principle of the assay The kit assay Human NRG-1 level in the sample，use Purified NRG-1 antibody to coat microtiter plate wells, make solid-phase antibody, then add NRG-1 to wells, Combined NRG-1 antibody which With HRP labeled goat anti-Human 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. The concentration of Human NRG-1 in the samples is then determined by comparing the O.D. of the samples to the standard curve. Materials provided with the kit 1 wash solution 20ml×1bottle 7 Stopp Solution 6ml×1 bottle 2 HRP-Conjugate reagent 6ml×1 bottle 8 Standard（） 0.5ml×1 bottle 3 Microelisa stripplate 12well×strips 9 Standard diluent 1.5ml×1bottle 4 Sample diluent 6ml×1 bottle 10 Instruction 1 5 Chromogen Solution A 6ml×1 bottle