![]() Towbin with or without SDS, CAPS, carbonate, Bjerrum Schafer-Nielsen Compatible Transfer Buffer by Gel Type Gel Type In addition, alcohol (methanol or ethanol) may be included in the transfer buffer to promote binding of proteins to membranes, and SDS may be added to promote elution of proteins from gels. Transfer buffers contain a conductive, strong buffering agent (for example, Tris, CAPS, or carbonate) in order to maintain the conductivity and pH of the system during transfer. Preinstalled, customizable programs for transfers of most proteins, user-programmable settings for traditional semi-dry techniquesĬooling with ice pack or refrigerated water recirculator ![]() Power and transfer time limited due to lack of cooling options Widest range of power settings and transfer times Comparison of Western Blotting Transfer Systems Filter papers and membranes are pre-wet and packaged in single-use packages, simplifying assembly of the transfer stack. Transfer conditions must be optimized for proteins of different molecular weights, e.g., to prevent under-transfer (incomplete transfer of proteins out of the gel) or over-transfer (loss of proteins passing completely through the membrane).Ī recent development, these systems utilize specialized apparatus that use proprietary filter papers and specialized buffers to rapidly and efficiently transfer proteins. Both the voltage and the distance between the electrodes then play a major role in governing the rate of elution of the proteins from the gel. The electric field strength (measured in V/cm) that is generated between the electrodes is the driving force for transfer. Voltage is applied between the electrodes and proteins migrate to the membrane following the current that is generated by the applied voltage across the electrodes. During this process, the membrane and gel are placed together, with filter paper between two electrodes. The most common method of transfer in western blotting is electrophoretic transfer, where an electric field is used to elute proteins from gels and transfer them to membranes. Gel and Membrane Setup for Electrophoretic Transfer ![]()
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