For the isolation of very high yields of 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/Ni 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/Ni products consists of crosslinked and beaded 6 % agarose, IDA-conjugated and charged with divalent Nickel ions. Nickel chelates recognize two exposed histidines with good specificity and very high affinity, making the Ni2+ charged matrix the first choice for all standard applications. ROTI®Garose His/Ni beads give very high yields of pure His-tagged protein 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).
A special feature is the modified polychelate linker used in the ROTI®Garose-His/Ni HPBeads plus (0806). For more detailed data please see this product.
For insulation under reducing conditions we recommend ROTI®Garose-His/Ni HPBeads plus or 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/Ni HPBeads, the advantages of the highly efficient Nickel-His binding have been combined with a bead technology allowing a very rapid flow rate. ROTI®Garose-His/Ni HPBeads are ideal for purifying large sample volumes.
50 % bead suspension in 20 % ethanol.
MPLC and FPLC, or if 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 | nickel |
Loading capacity (gel) (Ni2+) | ~20 µmol/ml |
Size exclusion limit (Da) | ~4x106 |
Ethanol | 20 % |