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 histidines that are in close proximity and accessible 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 agarose beads for affinity chromatography under presence of EDTA or DTT. The matrix of choice for applications under low or medium pressure, and applicable for every required sample and/or matrix volume.In ROTI^®Garose-His/Ni HPBeads plus, the advantages of the highly efficient Nickel-His binding have been combined with a bead technology allowing a very rapid flow rate. The specially developed polychelating ligand forms highly stabile nickel chelates, which are able to isolate His-tagged proteins even from solutions containing 20 mM EDTA or DTT - reliably and without undesired nickel stripping.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 from reducing solutions.

In the matrix of ROTI^®Garose-His/Ni HPBeads plus, IDA was replaced by a polychelator conjugation. This multi dentate cross-linker binds his-tagged proteins very efficiently, leading to high recovery rates with minimized nickel bleeding into the eluate. ROTI^®Garose-His/Ni HPBeads plus may not be regenerated.The matrix is very stabile and may generally be used with buffers including the following reagents: ≤20 mM EDTA, ≤20 mM DTT, ≤8 M urea, ≤6 M guanidinium hydrochloride, ≤100 % methanol, ≤100 % ethanol, ≤30 % acetonitrile, pH range 4-9. May be autoclaved at 121 °C for 30 mins.