Tris-Glycine Transfer Buffer (25X)

For non-sequencing applications, any transfer buffer used with Tris-Glycine gels can be used with Tricine gels including Tris-Glycine transfer buffer. For sequencing applications, the buffer should be chemically compatible with sequencing protocols. Non-glycine based transfer buffers such as the NuPAGE Transfer buffer, 1/2X TBE Transfer buffer, or CAPS Buffer can be used for N-terminal sequencing . Generally, a pH which is close to neutral is desirable to maintain gel and protein stability. High current should be avoided because it can lead to heat generation and instability.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

This is perfectly acceptable with the XCell II Blot Module. The liquid in the outer buffer chamber only serves as a coolant or heat sink. The reason why water is recommended is because it is a less expensive alternative.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

We recommend discarding the buffer and remaking it after rechecking the reagents and the water purity. We do not recommend adjusting the pH with acid or base, as this will increase the conductivity of the buffer and result in higher current during the transfer.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.