PureCube Ni-IDA Agarose
The His tag is the most widely used affinity tag due to its small size and versatility under native and denaturing conditions, as well as in presence of detergents and many other additives. Cube Biotech offers high-performance PureCube Ni-IDA Agarose, based on BioWorks Workbeads, for purification of his-tagged proteins. To detect His-tagged proteins in Western Blot experiments, Cube Biotech offers tag-specific antibodies
. For increased specificity, we offer a range of different His affinity resins, e.g. carrying the NTA ligand (what's the difference?
). Please see our overview page for more information.
Why PureCube Ni-IDA Agarose?
Binding capacity of up to 50 mg/mL
Stable under commonly used concentrations of DTT and EDTA
Can be regenerated for reuse
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 2 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-NTA Agarose and two equivalent market-leading resins.
| ||Cube Biotech||Competitor G||Competitor S|
||average 90 µm
|Metal ion capacity (Cu, Ni)
||> 40 mg/ml
|pH stability (processing)
|Recommended flow rate
||0.5-2.0 mL/min (6.0 mL/min poss.)
Table 1. Technical and performance parameters of 3 Ni-IDA agarose 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.