Can TALON columns/resin be regenerated?

Yes, TALON columns can be regenerated by washing with a MES/NaCl buffer to remove the imidazole. However, it cannot be guaranteed that the binding capacity after regeneration will be the same as that of the original unused resin. Takara Bio recommends that a regenerated column only be used to purify the same protein that it was previously used to purify, to prevent any contamination. Follow the [...]

Why does the column turn violet upon addition of the sample/buffer containing guanidine-HCl?

Violet color in the guanidine-HCl buffer is normal. It is caused by a weak coordination of the guanidine to the metal, causing a change in the spin energy of the d-orbital electrons that give cobalt its pink color, which slightly alters the hue of the metal ion’s color. This does not affect the binding capacity or the affinity characteristics of the column material. Imidazole binding to the [...]

If the HisTALON Gravity Column provides 1 ml of space for the resin slurry, how much extra room is there for the sample?

HisTALON Gravity Columns contain 1 ml of resin and have enough remaining space to hold another 6 ml of buffer without the cap, or 5 ml of buffer or sample with the cap on the column. The gravity columns are also available empty, without resin, as the TALON 2 ml Disposable Gravity Column (Cat. # 635606).Protein purification [...]

What is the chelator present in TALON resin?

The tetradentate chelator present in the core of TALON resin is a patented carboxymethyl aspartate.Protein purification [...]

What is metal ion transfer?

Intermediate metal ions (such as Cu2+, Ni2+, and Co2+) will bind to thiol groups. Dithiothreitol (DTT) can extract the metal ion from the resin, which is referred to as metal ion transfer. This also happens at high beta-mercaptoethanol (BME) concentrations. When this occurs, the resin will turn brown and may elute a brown precipitate in the flowthrough or washes. However, Takara Bio's studies [...]