ProtoBlot Rapid Western Blotting Buffer (10X)

ProtoBlot Rapid Western Blotting Buffer is a relatively low conductivity buffer designed to replace tris/glycine (Towbin) buffer in wet (tank) transfer applications. It allows protein transfer from gels to nitrocellulose or PVDF at relatively higher voltages.

The speed of transfer of a protein from the gel to the membrane during a western blot is proportional to the electric field strength. It is directly proportional to the voltage applied and inversely proportional to the distance between the electrodes and is expressed in V/cm.  The amount of heat produced during the transfer is a function of the current flowing through the system. A lower conductivity buffer like ProtoBlot can be run at a higher voltage producing less heat because the current conducted will be lower. This allows for a shorter run time and more efficient transfer.

Key notes:

• For use the 10X buffer is diluted to 1X and ethanol or methanol is added to 10%-20%.
• The pH of the buffer at a 1X dilution is 8.4–8.5 which is the ideal pH for maximum transfer of most proteins.
• The transfer can be done with or without an ice pack or cooling system.
• Optimum transfer times will vary and have to be determined empirically, but a suggested starting point is 300V for 20 minutes.