小鼠肝配蛋白B2(EFNB2) elisa试剂盒的注意事项：

1） 充分混匀对保证反应结果很重要，在加液后请轻轻晃动整个96板孔，以保证混匀。

2） TMB溶液请勿接触氧化剂或金属，否则容易失效。

3） 加样时请注意每个样品或标准品必须更换枪头，一方面避免交叉污染，另一方面也避免吸取体积的误差。加样过程中必须避免气泡的产生。

4） 检测标准品和样品时建议设置重复孔，以确保检测结果的可信度。

5）洗涤过程非常重要，洗涤不充分会下降并导致结果误差较大

Assay procedure：1.  Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.2.  Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.3.  Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank well doesn’t add anyting.4.  Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C.5.  Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6.  Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7.  Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8.  Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.