超薄切片(100nm) 0.01MPBSpH7.4(1%BSA+0.05%Tritonx-100 +0.05%Tween 20)封闭 倒扣5min,RT
一抗0.01MPBSpH7.4(1%BSA+0.05%Tween 20))稀释100-300倍,RT孵育2h或 4℃过夜
洗涤6x2min,RT 0.01MPBSpH7.4
二抗 0.01MPBSpH7.4 ( 1%BSA+0.05%Tween 20 ) )稀释 50倍,RT孵育1h
6x2min 0.01MPBSpH7.4 洗涤 , RT
0.01ddH2OpH7.4 洗涤 6x2min , RT
镍网干燥后双染色

