semi-dry transfer vs wet transfer
Posted 13 February 2004 - 10:20 PM
Posted 16 February 2004 - 04:12 AM
We're using both methods in our lab, so maybe I can give you some advice, though i can only tell you the difference between biorad's tank blots and biometra's semy-dry...
Each method has it's own pros and cons.
Semidry blotting is faster and easier, and reqires a lot less buffer volumes than tank blotting. Small Proteins (<20kDa) are transferred with better reproducibilty. Big proteins, however, do not transfer well in semidry blotting, and blotting of those is random at best, i.e. in some semidry blots you get big proteins on the membrane, sometimes you don't (same conditions!).
Tank blots transfer high molecular weight proteins (>60kDa) better than semi-dry, but more often than not some small proteins are blotted trough the membrane and are therefor lost for detection.
For the proteins I work with (25-35kDa) both methods work equally well, but I personally prefer the semi-dry blot, since I don't like wading though knee-deep in blotting buffer and preparing fresh buffer every other day. But for detection of BIG proteins (130kDa) I change to tank blots....
Posted 17 February 2004 - 02:22 AM
I also have doubt on another issue. That's protein transfer through the membrane and reach filter paper...lost of target protein!!
Both semidry and tank blot encounter this problem??
I've experienced that large protein (the marker shpwed that) transfer through the membrane! Will tank blot also have this problem?? Also large protein???
Posted 17 February 2004 - 07:59 AM
Yes, blotting through the membrane can happen with both systems....
If that's your problem, try to shorten the blotting time and/or lower the used amperage/voltage... Do some test blots with your standard and observe when at which conditions the transfer for a given size of protein is best...
Posted 24 August 2004 - 01:40 AM
Posted 15 September 2004 - 12:09 PM
Posted 09 May 2012 - 05:08 AM
A great devices to transfer protein is the Invitrogen iBlot mahine, transfer is done efficiently in 8mn and probing gives excellent results (we are using it daily)
Posted 09 May 2012 - 07:50 AM
genius does what it must
i do what i get paid to do
Posted 15 May 2012 - 06:36 AM