semi-dry transfer for midi gels - (Aug/23/2007 )

Hi everybody,


I was wondering if any of you had any experience with the Hoefer (Amersham/GE) semi-dry transfer unit. According to the company's user manual you can transfer 2 18X16 cm gels if you stack 2 sandwiches one on top of the other seperated by cellophane paper. Has anyone ever tried doing this and can tell me if the transfer effiency is the same for both of the gels? Another thing I was wondering about is the effiency for proteins with a high molecular weight - is it possible to use the semi-dry system for this and just run it for more time as I would if I was using the wet system?


Thanks,

Ilana

Hi everybody,


I was wondering if any of you had any experience with the Hoefer (Amersham/GE) semi-dry transfer unit. According to the company's user manual you can transfer 2 18X16 cm gels if you stack 2 sandwiches one on top of the other seperated by cellophane paper. Has anyone ever tried doing this and can tell me if the transfer effiency is the same for both of the gels? Another thing I was wondering about is the effiency for proteins with a high molecular weight - is it possible to use the semi-dry system for this and just run it for more time as I would if I was using the wet system?


Thanks,

Ilana

when you transfer 2 (or more) gels in semi-dry, they need to be very well matched (in size) and overlaid very carefully. the "cellophane" referred to is a porous cellulose membrane (dialysis sheet).

high mw proteins transfer poorly in semi-dry (there are some things you can do to improve transfer efficiency). you can increase transfer time but you have to be careful that the filter papers don't dry out during the transfer.

Thanks for the replies,

I think I'll just go with the wet system and save myself the trouble

while we're on the subject of blotting high mw proteins - I have a protein that's 480 KDa -

Does anyone have any advice on the process of detecting such a protein (gel %, transfer time, buffers, etc..)?
I've read somewhere that it's not recommended to use a standard Tris-glycine gel.

If anyone has any experience with this - any advice will be appreciated

Thanks for the replies,

I think I'll just go with the wet system and save myself the trouble

while we're on the subject of blotting high mw proteins - I have a protein that's 480 KDa -

Does anyone have any advice on the process of detecting such a protein (gel %, transfer time, buffers, etc..)?
I've read somewhere that it's not recommended to use a standard Tris-glycine gel.

If anyone has any experience with this - any advice will be appreciated