Nickel nitrilotriacetic acid

Manufactured by Thermo Fisher Scientific

Nickel-nitrilotriacetic acid (Ni-NTA) is a metal-chelating compound used in affinity chromatography for the purification of recombinant proteins containing a histidine (His) tag. It binds to the His-tag with high specificity, allowing the target protein to be isolated from complex mixtures.

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Lab products found in correlation

Ni nta agarose, by Thermo Fisher Scientific (1 mentions) Expi293 expression system, by Thermo Fisher Scientific (1 mentions) Akta purifier, by GE Healthcare (1 mentions) Frac 950 collector, by GE Healthcare (1 mentions) Microbca, by Thermo Fisher Scientific (1 mentions) Pet28a vector, by GenScript (1 mentions)

Close spelling variants of this product

Nickel nitrilotriacetic acid (Ni–NTA) beads HisPur nickel-nitrilotriacetic acid (Ni-NTA) resin Nickel-nitrilotriacetic acid agarose resin HisPur nickel-nitrilotriacetic acid (Ni-NTA) agarose Nickel-nitrilotriacetic acid resin HisPur™ Nickel-nitrilotriacetic acid-agarose (Ni-NTA) resin Nickel-nitrilotriacetic acid-agarose Nickel-nitrilotriacetic acid (Ni-NTA) agarose resin Nickel-nitrilotriacetic acid (Ni-NTA) resin

2 protocols using nickel nitrilotriacetic acid

Production and Purification of SARS-CoV-2 RBD

Cited 1 time

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SARS-CoV-2 spike receptor binding domain protein (RBD) was produced by transient transfection of HEK-293F cells with plasmid pCAGGS (32 (link), 33 (link)), containing the SARS-CoV-2 Wuhan-Hu-1 spike glycoprotein gene RBD with C-terminal hexahistidine tag (generously provided by F. Krammer). Transfection was carried utilizing the Expi293 expression system (Gibco, Thermo Fisher Scientific) following the manufacturer’s instructions. RBD was purified by affinity chromatography utilizing nickel-nitrilotriacetic acid (Ni-NTA) agarose (Thermo Fisher Scientific), and RBD purity was assessed by SDS-PAGE.

Gunawardana M., Breslin J., Cortez JM J.r., Rivera S., Webster S., Ibarrondo F.J., Yang O.O., Pyles R.B., Ramirez C.M., Adler A.P., Anton P.A, & Baum M.M. (2021). Longitudinal COVID-19 Surveillance and Characterization in the Workplace with Public Health and Diagnostic Endpoints. mSphere, 6(4), e00542-21.

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Synthesis and Purification of Azide-Tagged IFN-γ

Cited 1 time

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We synthesized azide-tagged IFN-γ (azIFN-γ) as previously reported,^{9 (link)} with no deviations. Briefly, azIFN-7 was inserted into a pET28a vector (GenScript), while N-myristoyltransferase was inserted into a pET11c vector (generously provided by Dr. Edward Tate, Imperial College London²⁴). We co-expressed both proteins in BL21 (DE3) E. coli, using 12-azidododecanoic acid (synthesized as in^{9 (link)}) as the azide tag, isolated the protein under denaturing conditions via nickel-nitrilotriacetic acid (Thermo Scientific), refolded it, and performed the final purification using fast protein liquid chromatography (FPLC, GE AKTApurifier 10 with Frac-950 collector - buffers in,^{9 (link)} procedure in^{25 (link)}). We determined concentration for all experiments using the bicinchoninic acid (Micro BCA, Thermo Scientific) assay.

Ham T.R., Cox D.G, & Leipzig N.D. (2019). Concurrent delivery of soluble and immobilized proteins to recruit and differentiate neural stem cells. Biomacromolecules, 20(9), 3445-3452.

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