You may get some amount of your protein transferred. For control of effectiveness, you can stain your membrane with a reversible protein stain like Ponceau S or some similar alternatives (e..g. https://www.lifetechnologies.com/order/catalog/product/24580 ). You can also counterstain your gel with EZBlue ( http://www.sigmaaldrich.com/life-science/proteomics/protein-electrophoresis/coomassie-stains.html ) after blotting. This may give you an idea of how much of the high-molecular weight proteins have been transferred.

If you exchange nitrocellulose for PVDF, longer transfer times are also acceptable. PVDF binds proteins much better than nitrocellulose.

Hi there,

The wet transfer condition you mentioned (I guess it's wet transfer) seems adapted for a classical overnight transfer according to the Biorad documentation (see attached document page 8). If I were you I would try different conditions and, as Christoph suggested, use Ponceau staining to quickly state on transfer efficiency. Knowing that HMW proteins are naturally poorly transferred, I guess you will have to optimize (gel composition, buffer composition aso)... The attached document also gives lines for improving transfer efficiency. Hope it helps.

Hello!

We use 300mA transfer for 2-3h for proteins with the mass more than 100kDa, and for this experiments I agree with Christoph on using PDVF.

The iBlot system can work quite well on large ~150kDa proteins in about 10 min, while smaller proteins transfer very well at 6 minutes. The key is high current, as it uses 25 VDC and 5.5 Amp. Complex membrane proteins can have more involved buffer requirements but most proteins work quite well in this "dry" transfer system.

Hello

I have performed a wet transfer using nitrocellulose membrane for high molecular weight protein up to 200KDa between 1 and half to 2 hours at 300mA, on Ice and it works well for me. Just ensure you use the right percentage gel for your protein separation when you perform your gel electrophoresis.