Can nonphosphate-based buffer systems such as Tris or acetate buffers be used with TALON resin?
Can nonphosphate-based buffer systems such as Tris or acetate buffers be used with TALON resin?
No, Tris or acetate buffers are not recommended for use with TALON resin. The correct pH cannot be obtained with sodium acetate, so it will interfere with binding. However it can be used for a pH-based elution buffer. Beyond this, an acetate buffer system is more conducive to microbial growth and is therefore less desirable. Using Tris buffers above 50 mM is not advised because Tris contains primary amines that may also interfere with his-tag affinity. Therefore, sodium phosphate is recommended. Furthermore, thiocyanate is not compatible with TALON resin. MOPS buffers perform slightly better than Tris buffers since the amine is not a primary amine, and HEPES buffer systems are known to work as well. Again, consult the TALON Metal Affinity Resins User Manual for a complete listing of compatible reagents.