Did you pre-sock and activate your PVDF with Methanol? Check your transfer buffers? Were they made correctly? We use: 25 mA per membrane and maximum voltage set at 35V for 0.5 hour. It transferred really well.
Yes, I activated my PVDF with Methanol for 5 min. I will try to set up 25 MA next time. For voltage, since the protocol said: Do not exceed 25 volts, so I never try higher voltage.
【在 q*****n 的大作中提到】 : Did you pre-sock and activate your PVDF with Methanol? : Check your transfer buffers? Were they made correctly? : We use: : 25 mA per membrane and maximum voltage set at 35V for 0.5 hour. : It transferred really well.
d*o
23 楼
上面没有刻你的mitbbs id,不能证明是你的设计
A*g
24 楼
下次会延长时间。 我一般也就是设定电压,但发现转膜过程中,电流会逐渐降低,会降到很低,就以为时 间延长没有什么用。 还有,再请教下,跟滤纸会不会也有关系呢,我用了Biorad的Extra thick blot paper 滤纸。
i compared the result side by side for a 300kd protein transfer. traditional wet transfer 1hr vs. iblot 7+1 additional minutes conclusion: traditional way beat iblot by at least 5 fold.