No. of hours recommended for blotting

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medfreak
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No. of hours recommended for blotting

 Hey guys,

So I was about to start the blotting of a gel (that has two portions: 12 % and 8 %).
And what I'm studying is muscle proteins (specifically: a mitochondrial enzyme: cytochrome oxidase).

I've set the voltage at 35 volts. Now: do you think transferring for 6 hours is a good idea?

medfreak
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 Anyone? 

 Anyone? 

protoldo
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Hola,  By the voltage choosen

Hola,  By the voltage choosen for you I think that you want transfer in a liquid immerse device. But when I worked with it , 1hour at 120 volts was enought. At least for proteins with mol. weight under 100kD. Now, I use the semidry transfer device and with 25volt, 25 min is enough to see a 75 kD protein and the partially the 180 marker in a 12% gel. Moreover since I use the semidry method there isn´t strong difference between the amount of protein transferred of high mol. wheight. and the low  M.W. ones, wich in the inmersion system was more evident . So with the risk of lost a part of the signal i only run a gel which after transfer I stain with c.blue .  Good luck

medfreak
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 Thanks a lot.

 Thanks a lot.