14,15-环氧化二十碳三烯酸(14,15-EET/DHET

（共享）ELISA原理与实验方法

.14 Just to check enzyme and color product put 100 µl of each in a test tube and watch for a yellow color change.Figure 1. Schematic of an ELISAMATERIALSPhosphate-buffered saline (PBS) ,0.01 M Na2HPO4,0.15 M MaCl, PH 7.4Sodium phosphate dibasic (Na2HPO

（交流）细胞ELISA

Cellular ELISA ProtocolFormalin Fixed Cell Plates1. Trypsinize confluent flasks2. Pool and count cells3. Centrifuge at 1500 rpm for 10 minutes4. Resuspend to the appropriate concentration in complete medium4 x 105 cells/ml for epithelial cells2 x

捕获ELISA法

PBST（0.05％Tween-20）洗涤孔盘三次2.13加入第二抗体的方式为加入HRP结合的山羊抗兔IgG（H + L）于适当的孔中。2.14孵育孔盘于室温下1小时。2.15用PBST（0.05％Tween-20）洗涤孔盘五次，。2.16加入入150微升底物（OPD，400微克/毫升，0.03％过氧化氢，柠檬酸盐缓冲液）2.17在室温下孵育30分钟。2.18读取450nm的吸光度。