For the isolation of His-tagged proteins under 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 matrix of ROTI®Garose-His/Ni NTA products consists of crosslinked and beaded 6 % agarose, NTA-conjugated and charged with divalent Nickel ions. Nickel chelates recognize two exposed histidine tags with good specificity and very high affinity, making the Ni2+ charged matrix the first choice for all standard applications. The tetradentate NTA cross-linker binds his-tagged proteins very efficiently, leading to high recovery rates with minimized nickel bleeding into the eluate.
ROTI®Garose-His/Ni NTA-Beads may 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) and (dependent on the respective buffer) reducing substances (for instance ≤30 mM glutathion, ≤10 mM DTT, ≤10 mM DTE, ≤20 mM β-mercaptoethanol und ≤0,3 % SDS).
Nickel charged NTA-agarose beads for affinity chromatography under reducing conditions. The matrix of choice for applications under medium pressure, with high flow rate or with big sample-/matrix volume.
In ROTI®Garose-His/Ni NTA-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 from reducing solutions.
May be autoclaved at 121 °C for 30 mins.
Gebruikstip | Bead suspension, binding capacity ≥60 mg/ml packed matrix (His)6 |
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Particle size | 50-150 µm |
Beads (cross-linked agarose) | 6 % |
Ligand | NTA |
Metal ion | nickel |
Binding capacity (gel) (GFPuv-6xHis) | ≥60 mg/ml |
Ethanol | 20 % |