NTA succinimidyl ester

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Telephone	1-800-990-8053
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Physical properties

Molecular weight	515.52
Solvent	DMSO

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

Alternative formats

Overview SDS Protocol

Molecular weight

515.52

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) is such a ligand commonly used in commercially available resins for purifying proteins. On the other hand, tag-NTA conjugates are used for detecting polyhistidine-tagged proteins. NTA succinimidyl ester is an excellent building block that can be readily used to attach NTA groups to a biological substrate or a surface that has amino groups. Through the NTA moiety attached to the substrate or surface, genetically expressed proteins bearing the hexahistidine extension (HIS Tag) can be detected or immobilized via Ni-mediated His-Tag method.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of NTA succinimidyl ester to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	193.979 µL	969.894 µL	1.94 mL	9.699 mL	19.398 mL
5 mM	38.796 µL	193.979 µL	387.958 µL	1.94 mL	3.88 mL
10 mM	19.398 µL	96.989 µL	193.979 µL	969.894 µL	1.94 mL

Molarity calculator

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Images

Figure 1. Chemical structure for NTA succinimidyl ester.

Citations

View all 1 citations: Citation Explorer

Transition Metal Ion FRET-Based Probe to Study Cu (II)-Mediated Amyloid-$\beta$ Ligand Binding
Authors: Wu, Ri and Svingou, Despoina and Metternich, Jonas B and Benzenberg, Lukas R and Zenobi, Renato
Journal: Journal of the American Chemical Society (2024)

References

View all 31 references: Citation Explorer

Proteomics analysis of host cell proteins after immobilized metal affinity chromatography: Influence of ligand and metal ions.
Authors: Lingg, Nico and Öhlknecht, Christoph and Fischer, Andreas and Mozgovicz, Markus and Scharl, Theresa and Oostenbrink, Chris and Jungbauer, Alois
Journal: Journal of chromatography. A (2020): 461649

Probing Protein-Protein Interactions with Label-Free Mass Spectrometry Quantification in Combination with Affinity Purification by Spin-Tip Affinity Columns.
Authors: Liu, Guizhen and Fu, Tao and Han, Ying and Hu, Shichen and Zhang, Xuepei and Zheng, Mengmeng and Hao, Piliang and Pan, Lifeng and Kang, Jingwu
Journal: Analytical chemistry (2020): 3913-3922

Promiscuity of host cell proteins in the purification of histidine tagged recombinant xylanase A by IMAC procedures: A case study with a Ni2+-tacn-based IMAC system.
Authors: Yang, Yuanzhong and Mitri, Khosse and Zhang, Chunfang and Boysen, Reinhard I and Hearn, Milton T W
Journal: Protein expression and purification (2019): 51-61

Detection and quantification of transition metal leaching in metal affinity chromatography with hydroxynaphthol blue.
Authors: Kokhan, Oleksandr and Marzolf, Daniel R
Journal: Analytical biochemistry (2019): 113347

Complementary IMAC enrichment methods for HLA-associated phosphopeptide identification by mass spectrometry.
Authors: Abelin, Jennifer G and Trantham, Paisley D and Penny, Sarah A and Patterson, Andrea M and Ward, Stephen T and Hildebrand, William H and Cobbold, Mark and Bai, Dina L and Shabanowitz, Jeffrey and Hunt, Donald F
Journal: Nature protocols (2015): 1308-18

Production of fully assembled and active Aquifex aeolicus F1FO ATP synthase in Escherichia coli.
Authors: Zhang, Chunli and Allegretti, Matteo and Vonck, Janet and Langer, Julian D and Marcia, Marco and Peng, Guohong and Michel, Hartmut
Journal: Biochimica et biophysica acta (2014): 34-40

Thermodynamics of imidazole-ligand binding to Ni-nitrilotriacetate in solution and covalently attached to agarose beads: imidazole, his-6 (his-tag) peptide and a new bis-imidazolo-dithiane.
Authors: Kirk, William R
Journal: Protein expression and purification (2014): 1-7

Quantitative analysis of global phosphorylation changes with high-resolution tandem mass spectrometry and stable isotopic labeling.
Authors: Kweon, Hye Kyong and Andrews, Philip C
Journal: Methods (San Diego, Calif.) (2013): 251-9

Development of recombinant single-chain variable fragment against hepatitis A virus and its use in quantification of hepatitis A antigen.
Authors: Nimmagadda, Sridevi V and Aavula, Shukra M and Biradhar, Neelakantam and Sula, Samuel and Lingala, Rajendra and Chandran, Dev and Villuppanoor, Srinivasan Alwar
Journal: Biologicals : journal of the International Association of Biological Standardization (2012): 299-308

Formation of high-capacity protein-adsorbing membranes through simple adsorption of poly(acrylic acid)-containing films at low pH.
Authors: Bhattacharjee, Somnath and Dong, Jinlan and Ma, Yiding and Hovde, Stacy and Geiger, James H and Baker, Gregory L and Bruening, Merlin L
Journal: Langmuir : the ACS journal of surfaces and colloids (2012): 6885-92