Immobilized Metal Ion Affinity Chromatography (IMAC), developed by Porath (1975), is based on the interaction of certain protein residues (histidines, cysteines, and to some extent tryptophans) with cations of transition metals. G-Sep™ Co IDA Agarose Fast Flow is specifically designed for the purification of recombinant proteins fused to the 6x histidine (6XHis) tag. Both nickel and cobalt IDA are offered as nickel has a higher binding affinity, but lower specificity compared to cobalt. This means that although cobalt IDA has a slightly lower efficiency compared to nickel IDA resin there is a significant reduction in non-specific binding.
The resin is specifically designed for the purification of recombinant proteins fused to the 6x histidine (6XHis) tag expressed in bacteria, insects, and mammalian cells. The resin is high affinity and selectivity for recombinant fusion proteins that are tagged with six tandem histidine residues.
G-Sep™ Co IDA Agarose Fast Flow (FF) resin is cobalt ions immobilized onto highly cross-linked 6% agarose beads using iminodiacetic acid groups (IDA). The G-Sep™ IDA Agarose Fast Flow (FF) resins have high chemical stability, allowing well proven cleaning-in-place (CIP) and sanitization protocols. G-Sep™ Ni IDA Agarose Fast Flow (FF) resin using nickel is also available
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- Matrix: Cross-linked agarose beads, 6%
- Bead form: Spherical, diameter 50-160μm
- Spacer: Epichlorohydrin
- Chelating Agent: Iminodiacetic acid
- Active group: Co2+
- Co2+ density: 20-40μmol /ml
- Binding Capacity: 5-10mg His-tagged protein/ml medium
- pH stability Working Range: 3-12
- pH stability Cleaning-in-Place (CIP): 2-14
- Maximum Flow Velocity: 450cm/h
- Exclusion limit(globular proteins): 4 x 106
- Physical Stability: Negligible volume variation due to changes in pH or ionic strength
- Chemical Stability: Stable to all commonly used aqueous buffers: 6M urea, 8M guanidine hydrochloride,
- Autoclavable: 121°C, pH 7, for 30 min
- Storage Conditions: 2 to 8°C, 20% Ethanol
- Immobilized meatl affinity chromatography (IMAC)
- Purification of 6X His tagged proteins
- Purification of cobalt binding proteins