For the isolation of high yields of highly pure His-tagged proteins under non-reducing conditions.
Immobilized metal ion affinity chromatography (IMAC) still is the most widely used method to obtain high yield of very pure proteins with reasonable effort.
ROTI®Garose His/Co Beads and ready-to-use columns offer the perfect solution for your specific applications in the field of benchtop column chromatography, for protein isolation in batch processes or under medium pressures (MPLC, FPLC). Depending on the product, our beads are optimally suited for low or high flow rates or when small or large sample volumes are to be purified. The ready-to-use columns and cartridges offer you the perfect solution for fast and easy purification of His-tag proteins from total lysate.
The matrix of ROTI®Garose His/Co products consists of crosslinked and beaded 6 % agarose, IDA-conjugated and charged with divalent Cobalt ions. Cobalt chelates recognize two exposed, vicinal histidines with superior specificity and good affinity, resulting in acceptable to good recovery rates only, but yielding proteins of superior purity. This matrix is, therefore, the perfect choice if extremely pure proteins are required, or if hard-to-separate proteins shall be isolated. ROTI®Garose His/Co beads result in eluates with considerably low metal contamination.
The tridentate IDA cross-linker provides easy elution with low amounts of imidazole. Additionally, ROTI®Garose Beads may repeatedly be regenerated, making them very cost-effective.
The Matrix is stable in all commonly used reagents including denaturing reagents (like 8 M urea, 6 M guanidinium hydrochloride, 5 mM DTT).
For insulation under reducing conditions we recommend ROTI®Garose-His/Ni NTA beads.
Nickel charged IDA-agarose beads for affinity chromatography under medium pressure, with high flow rate or with big sample-/matrix volume.
In ROTI®Garose-His/Co HPBeads, the advantages of the highly efficient cobalt-His binding have been combined with a bead technology allowing a very rapid flow rate. ROTI®Garose-His/Co HPBeads are ideal for purifying large sample volumes.
50 % bead suspension in 20 % ethanol.
MPLC and FPLC, or if superiorly pure His-tag proteins shall be isolated either in particularly short time or in big amounts.
May be autoclaved at 121 °C for 30 mins.
Gebruikstip | Bead suspension |
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Particle size | 50-150 µm |
Beads (cross-linked agarose) | 6 % |
Ligand | IDA |
Metal ion | cobalt |
Loading capacity (gel) (Co2+) | ~20 µmol/ml |
Size exclusion limit (Da) | ~4x10^6 |
Ethanol | 20 % |