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Wet transfer vs Semi-dry transfer
Posted by: sv112 (IP Hidden, New member, 1)
Date: June 4, 2006 12:32AM
Which transfer system (wet or semi-dry) does your lab prefer use and why? What are the advantages/disadvantages of each system? ***Thanks- your responses are greatly appreciated***
Edited 1 times. Last edit at 06/04/06 03:36AM by sv112.
Re: Wet transfer vs Semi-dry transfer
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: June 22, 2006 04:20PM
We use wet (tank) transfer, but both techniques are probably equally effective. Wet transfer is slow (at least one hour to overnight) and requires large volumes of buffer (~325ml-2L, depending on gel size). Semi-dry is far faster (15-45 minutes) as the buffer reservior is replaced with layers of filter paper soaked in buffer, which allows the plate electrodes to be in direct contact with the filter paper. However, most semi-dry methods require more than one type of buffer to achieve efficient transfer of both large and small proteins. In addition semi-dry blotting systems have lower buffer capacity, due to the huge reduction in buffer volume, and cannot be used for prolonged transfers. Once the capacity is exceeded the transfer will either stop or become uneven. Keep in mind too that if you use semi-dry transfer that the filter paper and membranes must be cut to the exact size of the gel to force the current to flow through the gel. If the filter paper is too large, the current will short-circuit through the overlapping filter paper around the edges of the gel.
Re: Wet transfer vs Semi-dry transfer
Posted by: ianbirmingham (IP Hidden, New member, 7)
Date: July 5, 2006 10:50AM
Semi-dry- transfers mini gels in about 30 minutes, wet transfers in 4hour- overnight. I used to prefer semidry for speed, but I have to say I now have reverted to wet transfers due to reliability- with semi-dry you need to be much more careful to remove bubbles of air between membrane and gel. For semi-dry blotting I would recommend PDVF membrane as smaller proteins can pass right through nitrocellulose when using semidry blotting. Ian.
Re: Wet transfer vs Semi-dry transfer
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: July 7, 2006 10:44AM
we also use PDVF for tank transfer (wet) as this membrane is superior to nitrocellulose in protein binding capacity (100-300ug/cm2 vs 80-100ug/cm2) and binding in the presence of SDS. Also PDVF can be used for rapid immunodetection, whereas nitrocellulose is unsuitable.
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