TA-Chelating FF is an immobilized metal affinity chromatography medium, which is made by covalently cross-linking ligand iminodiacetic acid (IDA) into TA-FF matrix. The separation of target proteins is accomplished based on the interaction of side-chain histidine, cysteine and tryptophan with transition metal ions (Cu2+，Co2+，Ni2+，Zn2+, etc.) immobilized on the medium.
The ligand of TA-Chelating FF media can provide three ligand sites to chelate with metal ions, and at the same time provide three ionic bonding sites to purify the target proteins with high affinity, while the same type of TA-IMAC FF media provides four ligand sites to chelate with metal ions and two ionic bonding sites to purify the target proteins, that is, the same ligand density and the same conditions of metal ions. That is to say, under the same ligand density and the same metal ion condition, TA-Chelating FF medium has a stronger affinity than TA-IMAC FF, and all the samples that cannot be adsorbed in TA-IMAC FF medium can be selected to be bound with TA-Chelating FF. However, at the same time, because of the additional metal ion chelating site, TA-IMAC FF medium has a stronger binding strength for metal ions, and it can also be compatible with the reducing agents DTT and β-mercaptoethanol. mercaptoethanol, therefore, the medium should be selected according to the protein characteristics when purifying recombinant histidine-tagged proteins.
Specifications:
| Product |
TA-Chelating FF |
| Appearance |
White slurry, forms visible layering upon standing |
| Agarose Concentration |
6% |
| Particle Size Range |
45–165 µm |
| Metal Chelating Capacity |
Cu2+: approx. 34 µmol/mL |
| Max. Pressure Tolerance |
0.3 MPa |
| Flow Rate Range |
>300 cm/h (TK-EC50, h=15cm, 0.1 MPa, 25 ℃) |
| Operation pH |
4–8.5 |
| Chemical Stability |
When metal ion are removed:
Stable in common aqueous solutions, 8M urea, 6M guanidine hydrochloride |
| pH Stability |
3–13 (operating), 2–14 (CIP, metal ion removed) |
| Recommended Flow Rate |
60–200 cm/h |
