| 1. | Western blot analysis we treated 200 fertilized eggs in a series of steps including lysing them in l0ul of lysis buffer ( 50mm tris - hcl ph 7 . 5 , 250mm nacl , 5mm edta , imm dtt , 0 . 1 % triton , 50mm sodium orthovanadate , looug / mg pmsf and tpck , 50ug / ml tlck , 1 ug / ml leupeptin pepstatin and aprotinin ) , frozing them below - 70 , and then thawing them at room temperature for 10 min
2 . westem印迹将处理组及对照组小鼠一细胞g2期受精卵各200个取出，转移到ep - pendoff管中， 5000甲m离心4分钟，弃去上清，加入ro川粉碎缓冲液，充分振荡混匀，在液氮中经过2一3次的冻融循环，迫使卵细胞裂解，加人等量的样品缓冲液沸水中煮5分钟，用于westem印迹分析。 |
