TALON reagent compatibility

Overview

TALON purification resin lets you prepare exceptionally pure his-tagged proteins from bacterial, mammalian, yeast, and baculovirus-infected cells, under native or denaturing conditions. TALON is an immobilized metal affinity chromatography (IMAC) resin charged with cobalt, which binds to his-tagged proteins with higher specificity than nickel-charged resins. As a result, TALON resin delivers his-tagged proteins of the highest purity. In addition, each cobalt ion is bound to the resin at four sites, resulting in low metal ion leakage.

Reagents compatible with TALON Resin
Reagent	Acceptable Concentration
Beta-mercaptoethanol^a	10 mM (with caution)
CHAPS^b	1% (with caution)
Ethanol^c	30%
Ethylene glycol	30%
HEPES	50 mM
Glycerol	20%
Guanidine hydrochloride^a	6 mM
Imidazole^d	200 mM at pH 7.0-8.0, for elution
KCl	500 mM
MES	20 mM
MOPS	50 mM
NaCl	1.0 M
NP-40	1%
SDS^b	1% with caution
Tris^e	50 mM
Triton-X 100	<1%
Urea	8 M

Use resin immediately after equilibrating with buffers containing these reagents. Otherwise, the resin will change color. Do not store resin in buffers containing these reagents.
Ionic detergents like CHAPS (3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate), SDS (sodium dodecyl sulfate), and sarkosyl are compatible up to 1%. However, due to their charged nature, you should anticipate interference with binding, even at low concentrations.
Ethanol may precipitate proteins, causing low yields and column clogging.
Imidazole cannot be used at concentrations higher than 5–10 mM for loading his-tagged proteins, because it competes with the histidine side chains (imidazole groups) for binding to the immobilized metal ions.
Tris coordinates weakly with metal ions, causing a decrease in capacity.

Reagents incompatible with TALON Resin
DTT (dithiothreitol) and DTE (dithioerythritol) NOTE: Using strong reducing agents will interfere with cobalt metal ion binding to the resin. EDTA (ethylenediaminetetraacetic acid), EGTA (ethylene glycol tetraacetic acid), and PEI NOTE: Although you can use EDTA at indicated points, it must be removed from the sample by gel filtration prior to applying the sample to TALON Resins.

Reagents incompatible with TALON Resin

These reagents are incompatible at any concentration:

DTT (dithiothreitol) and DTE (dithioerythritol)

NOTE: Using strong reducing agents will interfere with cobalt metal ion binding to the resin.

EDTA (ethylenediaminetetraacetic acid), EGTA (ethylene glycol tetraacetic acid), and PEI

NOTE: Although you can use EDTA at indicated points, it must be removed from the sample by gel filtration prior to applying the sample to TALON Resins.

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