Which reagents are compatible with TALON resin?
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 with caution. EDTA, EGTA, and PEI are chelating agents that strip the metal ligand from the column.
Acceptable concentration of Reagents with TALON Resin
Beta-mercaptoethanola: 10 mM (with caution)
CHAPSb: 1% (with caution)
Ethanol: 30%
Ethylene glycol: 30%
HEPES: 50 mM
Glycerol: 20%
Guanidine hydrochloridea: 6 mM
Imidazoled: 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: 1% with caution
Trise: 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
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.