Biotin-X IDA

Immobilized metal affinity chromatography (IMAC) is a popular method for protein purification, particularly for recombinant proteins fused to a polyhistidine-tag. Transition metal ions immobilized to a matrix through a chelating ligand interact with the polyhistidine-tag, effectively sequestering the fused protein from a sample. Nitrilotriacetic acid (NTA) and iminodiacetic acid (IDA) are two such ligands commonly used in commercially available resins. AAT Bioquest offer a variety of NTA building blocks for developing either NTA-based purification and detection. IDA is complimentary to NTA. The tridentate IDA ligand requires a lower imidazole concentration to elute protein than the tetradentate NTA. IDA is a smaller molecule which can be coupled to the matrix at a higher density resulting in a higher metal loading capacity. Biotin IDA is a bifunctional reagent that can be used to detect histidine-tagged proteins immobilized. Biotin IDA can be removed from the histidine-tagged protein at pH 4.8, allowing the blot to be reanalyzed with another probe.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
12630	1 mg	Price

References

View all 9 references: Citation Explorer

Synchronized purification and immobilization of his-tagged β-glucosidase via Fe3O4/PMG core/shell magnetic nanoparticles.

Authors:

Zhou, Yang and Yuan, Shaofei and Liu, Qian and Yan, Dandan and Wang, Yun and Gao, Li and Han, Juan and Shi, Haifeng

Journal:

Scientific reports (2017): 41741

Grafting iminodiacetic acid on silica nanoparticles for facilitated refolding of like-charged protein and its metal-chelate affinity purification.

Authors:

Liu, Hu and Dong, Xiaoyan and Sun, Yan

Journal:

Journal of chromatography. A (2016): 277-83

Uniform magnetic core/shell microspheres functionalized with Ni2+-iminodiacetic acid for one step purification and immobilization of his-tagged enzymes.

Authors:

Zhang, Yuting and Yang, Yongkun and Ma, Wanfu and Guo, Jia and Lin, Yao and Wang, Changchun

Journal:

ACS applied materials & interfaces (2013): 2626-33

Dual affinity method for plasmid DNA purification in aqueous two-phase systems.

Authors:

Barbosa, H S C and Hine, A V and Brocchini, S and Slater, N K H and Marcos, J C

Journal:

Journal of chromatography. A (2010): 1429-36

Evaluation of IDA-PEVA hollow fiber membrane metal ion affinity chromatography for purification of a histidine-tagged human proinsulin.

Authors:

de Aquino, Luciana Cristina Lins and de Sousa, Heloisa Ribeiro Tunes and Miranda, Everson Alves and Vilela, Luciano and Bueno, Sônia Maria Alves

Journal:

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2006): 68-76

Physical properties

Molecular weight	515.63
Solvent	Water

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

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Page updated on October 13, 2025