Ni-IDA is a abbreviation for a nickel ion that has been coupled to Iminodiacetic acid (IDA). In the field of protein purification it is used for His-tag purifications. It can be used either as a agarose resin for FPLC or as magnetic beads. Cube Biotech offers high quality Ni-IDA products for your IMAC assays.
Overview of Cube Biotech's Ni-IDA products for His-tag protein purification.
Features
| Usage | Specific binding and purification of 6x His-tagged proteins |
| Specifity | Affinity to His-tagged proteins |
| Binding capacity | >70 mg/mL |
| Chelator stability | Stable in buffer containing 10 mM DTT and 1 mM EDTA |
| Bead Ligand | Ni-IDA |
Ni-IDA products from Cube Biotech was successfully used in the following publications:
Our Ni-IDA Resins in detail
Fig. 1: The binding capacity of PureCube Ni-IDA Agarose is comparable to resins from a leading supplier. SDS-PAGE of CAT purified in gravity columns with PureCube Ni-IDA Agarose and Ni resin from Competitor G show elution fractions (E1-E5) with similar protein yields.
Strong and robust performance
Based on the same agarose matrix with high porosity and physical stability, PureCube Ni-IDA agarose exhibits a protein capacity of 50 mg protein per mL resin, which is competitive with that of products from market-leading providers. Figure 1 shows the SDS-PAGE analysis of 6-His chloramphenicoltransferase (CAT) expressed in E. coli and purified on gravity flow columns filled with PureCUbe Ni-IDA Agarose or Competitor G Ni-Sepharose High Performance resin. The amount of protein drawn down from the columns in 5 elution fractions was comparable for the two affinity resins (CL: cleared lysate; FT: flow-through; W1-2: wash fractions; E1-5: elution fractions). Table 1 compares additional parameters among PureCube Ni-IDA Agarose and two equivalent market-leading resins.
Top-quality materials, competitive performance
PureCube Ni-IDA Agarose is produced with a highly cross-linked agarose which is physically very stable and suitable for purification processes under low pressure (bulk purifications as well as packed in chromatography columns). Additionally, the high porosity and uniform size of the matrix allows for optimal protein interaction and high reproducibility between individual purification runs. Table 1 summarizes technical and performance parameters of PureCube Ni-IDA Agarose and 2 equivalent, market-leading resins.
Table 1. Comparision of three Ni-IDA manufacturers on the market.
| | Particle Size | Metal ion capacity | Binding capacity | pH stability | Recommended flow rate | DTT stability | EDTA stability |
| Cube Biotech | 32-60 µm | >25 µmol/mL/mL | 70 mg/mL | 3.0-13.0 | 0.5-2.0mL/min (6.0 mL/min possible) | 10 mM | 1.0 mM |
| Competitor G | average 90 µm | >30 µmol/mL | 15 mg/mL | 3.0-13.0 | 1.0 mL/min | No information | No information |
| Competitor S | 45-160 µm | 6-18 µmol/mL | 70 mg/mL | Not provided | 1.0 mL/min | 5 mM | Not recommended |
References
- Heinzler, R. et al. (2019). Toward Automated Enzymatic Glycan Synthesis in a Compartmented Flow Microreactor System. Advanced Synthesis & Catalysis. 10.1002/adsc.201900709.
