Voltage_question

[krekkzs]

Dear Everybody!

Do you know what is the optimal voltage/voltage range to blot a protein on PVDF memrane?
How many V/Cm2?

Or how is connection between the MW and V?
thanks for your answer,
Zsu

[kiki06]

Hello there welcome to the forums.

Usually the proteins are transferred at 30V (volts), however most people transfer to western blot membranes for 3 hours at 300 mA or overnight at 30 mA (constant current mA=milliamps).

Constant current gels sharper results both during transferring and electrophoresis for western blotting and sds-page gels.

If you find more information, please let me know about this.

[admin]

Hello! just found out that people run gels at 1.5 - 2 V/cm2.

Please double check this value though I think its right.

[krekkzs]

Dear All,
Thanks for your help,
Zsu

Originally Posted by krekkzs

Dear Everybody!

Do you know what is the optimal voltage/voltage range to blot a protein on PVDF memrane?
How many V/Cm2?

Or how is connection between the MW and V?
thanks for your answer,
Zsu

[admin]

no problem cheers

[trypsin]

I read that it is usual to transfer to western blot at 1-3V/cm2. I always transfer to wb at 75 mA (0,75mm gels) and 90-100 mA (1,0 mm mini gels) for 1 hour or 1hour 10min. My blots are very good. AND the proteins don't go through the PVDF-membrane so you don't care about 5 or 10 min. I use the semi dry transfer system.
I guess it can depend on your transfer system. i think the current in semi dry transfer isn't the same like in wet transfer. I've never used the wet transfer system.
bye, trypsin