b'Wet transfer (tank transfer)Tank transfer systems are the most commonly used transfer systems, with semi-dry and dry transfer techniquesAnode core (+)being an evolution of traditional wet tank transfer. In thisSponge padDirection of transfermethod, the gel is first equilibrated in transfer buffer. TheFilter papergel is then placed in the transfer sandwich (filter papergel Transfer membranemembranefilter paper), cushioned by pads and pressedGeltogether by a support grid. The supported gel sandwich isFilter paperplaced vertically in a tank between stainless-steel/platinumSponge padwire electrodes and immersed in transfer buffer (Figure 3).Cathode core ()Multiple gels may be electrotransferred in the standard field option, which is performed either at constant current (0.1 to 1 A) or voltage (5 to 30 V) for as little as 30 minutes to overnight. Transfers are typically performed with an ice pack or at 4C to mitigate the heat produced. Figure 3. Tank (wet) transfer apparatus for western blotting, with gelmembrane sandwich detail. Schematic showing the assembly In wet transfer, transfer efficiencies are better for lowerof a typical western blot apparatus with the position of the gel and transfer membrane and direction of protein transfer in relation to the molecular weight proteins than higher molecular weightelectrode position.proteins, with typical efficiencies of 80100% for proteins between 14 and 116 kDa [2]. The transfer efficiency improves with increased transfer time. However, with increasing time and the use of membranes with larger pore sizes (0.45 m), the risk of transferring the proteins completely through the membrane increases (also knownDid you knowas blow-through), especially for lower molecular weightIn many cases, ethanol can be substituted for methanol (30 kDa) proteins. in transfer buffers without impacting transfer efficiency. 7'