TA-Chelating HP is an immobilized metal affinity chromatography medium, which is made by covalently cross-linking ligand iminodiacetic acid (IDA) into TA-HP 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 HP 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 HP media provides four ligand sites to chelate with metal ions and two ionic bonding sites to purify the target proteins, that is to say, under the same density of ligands and the same metal ions condition. That is to say, under the same ligand density and the same metal ion conditions, the affinity of TA-Chelating HP medium is stronger than that of TA-IMAC HP, so samples that cannot be adsorbed by TA-IMAC HP medium can be selected to be bound by TA-Chelating HP, but at the same time, because of the additional metal ion chelating site, TA-IMAC HP medium has a stronger strength to bind metal ions, and it can also be compatible with the reducing agents DTT and β-mercaptoethanol, therefore, it is compatible with 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 HP |
| Appearance |
White slurry, forms visible layering upon standing |
| Agarose Concentration |
6% |
| Particle Size Range |
24–44 µm (avg. 34 µm) |
| Metal Chelating Capacity |
Cu2+: approx. 34 µmol/mL |
| Max. Pressure Tolerance |
0.3 MPa |
| 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 |
60–150 cm/h |
