ProtoBlot Rapid Western Blotting Buffer (10X)
- Blot in half the time versus conventional electroblotting buffers
- Very high efficiency transfer
- Optimal pH; less harsh on proteins
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.
- 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.