What is the maximum PMSF concentration that can be used with TALON resin?

PMSF is used in Takara Bio's ProteoGuard Protease Inhibitor Cocktail at a concentration of 1 mM in the 1X dilution. We do not recommend concentrations higher than 1 mM.Protein purification [...]

What is the maximum imidazole concentration that can be used with IMAC resins such as TALON resin?

The maximum recommended imidazole concentration is 250 mM. Concentrations higher than 250 mM can lead to leaching of cobalt ions from the resin.Protein purification [...]

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 [...]

Which reagents are compatible with TALON resin?

TALON resin is compatible with many standard laboratory reagents. However, four reagents NOT compatible with TALON include DTT (dithiothreitol), EDTA (ethylene diamine tetraacetic acid), EGTA (ethylene glycol-bis(beta-amino-ethyl ether)), and PEI (polyethyleneimine). DTT creates thiol-reducing complexes which make the resin unstable. Recent Takara Bio studies have shown that 1 mM DTT can be used [...]

When can the resin no longer be used?

The resin can no longer be used if it turns opaque/milky white, if it turns brown, if it loses its pink color, or if the binding capacity drops too low. These changes are usually due to using incompatible reagents on the column or regenerating the resin too many times. This can also happen if the entire protein precipitates onto the column. The researcher may want to remove the precipitated [...]