ELISA实验原理:  用纯化的抗体包被微孔板，制成固相载体，往包被抗体的微孔中依次加入标本或者标准品、生物素化的抗体、HRP-标记的亲和素，经过彻底洗涤后用底物显色。底物在过氧化物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目标蛋白呈正相关。用酶标仪在(450nm)波长下测定测定O.D.值(吸光度)，计算样品浓度。         ELISA Test principle: The microtiter plate provided in ELISA Kit has been precoated with an antibody specific to the target protein. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated specific polyclonal antibody preparation,and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well.        Only those wells that contain target protein, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of the samples is then determined by comparing the O.D. of the samples to the standard curve.

为了获得更理想的ELISA实验结果，请您 1、在ELISA实验过程中，请精确配制标准品及工作液A和B，以避免由于不准确稀释而造成的浓度误差； 2、在ELISA实验过程中，酶标板在温育时应加膜覆盖以避免液体蒸发； 3、在ELISA实验过程中，检测液A和检测液B，以及底物溶液在使用前，应置于37℃温育30分钟待用； 4、在ELISA实验过程中，洗板后应尽快进行下一步操作，避免酶标板长时间处于干燥状态。 In order to obtain the optimal ELISA results 1．During ELISA assay operation, Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction and avoid foaming and mix gently until the crystals have completely dissolved; 2．During ELISA assay operation, Proper adhesion of plate sealers during incubation steps is necessary; 3．During ELISA assay operation, Substrate, Detection Reagent A and Detection Reagent B should be incubate for 30 min at 37℃ before use; 4．During ELISA assay operation, After the every step of aspiration / wash, please do not delay entering into the next step in order not to let the assay plate "too dry".