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17 kDa ideal transfer voltage - PVDF


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4 replies to this topic

#1 LacquerHead

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Posted 15 November 2012 - 03:17 PM

Hi all,

Wondering what people would use ideally to transfer a 17kDa protein as efficiently as possibly onto PVDF? I have been doing 1.5 hours at 75V (constant voltage) but am slightly losing the low range. Open for some ideas here. Thanks!

#2 PhDinAcronyms

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Posted 15 November 2012 - 04:09 PM

Is that wet transfer? I transfer a 17kDa protein to PVDF by semi-dry at 23V for 1hr. Either way if you're losing the smaller proteins to blow through, then shorten the time or lower the voltage or increase methanol concentration.

#3 LacquerHead

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Posted 15 November 2012 - 04:30 PM

Yes wet transfer, I was going to try 75V for an hour? Should be ok I think for the smaller stuff.

#4 bob1

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Posted 15 November 2012 - 05:10 PM

75 V for 1 hour is enough to transfer big proteins (100 kDa) across, try about 30 V for an hour.

#5 mdfenko

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Posted 16 November 2012 - 11:04 AM

what is the pore size of the membrane? for low molecular weight proteins you should use 0.22um or smaller.

you can determine if you are blowing your protein through the membrane by using a backup membrane.
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