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Biotin-X IDA

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Physical properties
Molecular weight515.63
SolventWater
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Molecular weight
515.63
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.

Calculators


Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Biotin-X IDA to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM193.938 µL969.688 µL1.939 mL9.697 mL19.394 mL
5 mM38.788 µL193.938 µL387.875 µL1.939 mL3.879 mL
10 mM19.394 µL96.969 µL193.938 µL969.688 µL1.939 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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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
Overproduction of Thermus sp. Strain T2 beta-galactosidase in Escherichia coli and preparation by using tailor-made metal chelate supports.
Authors: Pessela, Benevides C C and Vian, Alejandro and Mateo, César and Fernández-Lafuente, Roberto and García, José L and Guisán, José M and Carrascosa, Alfonso V
Journal: Applied and environmental microbiology (2003): 1967-72
Evidence for phosphorylation of serine 753 in CFTR using a novel metal-ion affinity resin and matrix-assisted laser desorption mass spectrometry.
Authors: Neville, D C and Rozanas, C R and Price, E M and Gruis, D B and Verkman, A S and Townsend, R R
Journal: Protein science : a publication of the Protein Society (1997): 2436-45
One-step purification, covalent immobilization, and additional stabilization of a thermophilic poly-His-tagged beta-galactosidase from Thermus sp. strain T2 by using novel heterofunctional chelate-epoxy Sepabeads.
Authors: Pessela, Benevides C C and Mateo, Cesar and Carrascosa, Alfonso V and Vian, Alejandro and García, José L and Rivas, German and Alfonso, Carlos and Guisan, José M and Fernández-Lafuente, Roberto
Journal: Biomacromolecules : 107-13
Cell chromatography: separation of different microbial cells using IMAC supermacroporous monolithic columns.
Authors: Dainiak, Maria B and Plieva, Fatima M and Galaev, Igor Yu and Hatti-Kaul, Rajni and Mattiasson, Bo
Journal: Biotechnology progress : 644-9