b'Semi-dry transferSemi-dry transfer became available as the need for fasterMethanol may be included in the transfer buffer, but results became an issue for researchers. For semi-dryother organic solvents, including aromatic hydrocarbons, protein transfer, the transfer sandwich is placed horizontallychlorinated hydrocarbons, and acetone, should not between two plate electrodes in a semi-dry transferbe added to avoid damage to the electrode plates. apparatus (Figure 5). The key to improving the speedFast-blotting, semi-dry techniques use higher ionic of transfer with this method is to maximize the currentstrength transfer buffers and a high current power supply passing through the gel versus around it. To do this, theto decrease transfer times to under 10 minutes. In rapid amount of buffer used in the transfer is limited to thatmethods, amperage is held constant and voltage is limited contained in the transfer sandwich. Hence, it is critical thatto a maximum of 25 V. Transfer with traditional Towbin the membrane and filter paper sheets are cut to the gelbuffers can be preformed in a semi-dry apparatus either at size without overhang and that the gel and filter paper areconstant current (0.1 up to approximately 0.4 A) or voltage thoroughly equilibrated in transfer buffer. Also, the use of(10 to 25 V) for 30 to 60 minutes.extra-thick filter paper (approximately 3 mm thickness) is helpful in certain semi-dry transfer devices because these sheets can hold more transfer buffer.Did you knowAddition of SDS to the transfer buffer increases the 1 relative current, power, and heating during transfer, and may also affect the recognition of some proteins by antibodies.23451 Cathode2 Transfer gel layer (cathode)3 Gel4 Transfer gel layer (anode), membrane on top5 AnodeFigure 5. Semi-dry electroblotting transfer. The filter papergelmembranefilter paper sandwich is placed horizontally between two plate electrodes for semi-dry transfer. The presoaked filter paper acts as an ion reservoir that enables fast transfer.10'