Nickel nitrilotriaceticacid nta resin

Manufactured by Qiagen

Nickel-nitrilotriaceticacid (NTA) resin is a chromatographic resin designed for the purification of histidine-tagged recombinant proteins. The resin consists of nickel-charged nitrilotriacetic acid (Ni-NTA) immobilized on agarose beads. The histidine-tag on the target protein binds reversibly to the nickel ions, allowing for selective capture and elution of the protein of interest.

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Lab products found in correlation

Isopropyl β d thiogalactopyranoside, by Merck Group (1 mentions) Hiprep 16 60 sephacryl s 200 high resolution column, by GE Healthcare (1 mentions)

Close spelling variants of this product

Nickel-NTA resin Nickel-NTA Nickel resin NTA) resins

2 protocols using nickel nitrilotriaceticacid nta resin

Equine Alcohol Dehydrogenase Purification

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Primers were synthesized by Integrated DNA Technologies. Recombinant
alcohol dehydrogenase from Equus caballus (HLADH)
was obtained as described previously.^{30 (link)} Reduced β-nicotinamide adenine dinucleotide (NADH), isopropyl
β-d-thiogalactopyranoside (IPTG), and benzoylformate
were purchased from Sigma-Aldrich. A gel filtration calibration kit
(high molecular mass) and HiPrep 16/60 Sephacryl S-200 High Resolution
column were purchased from GE Healthcare. Nickel-nitrilotriacetic
acid (NTA) resin was purchased from Qiagen. Pfu DNA
polymerase was purchased from Stratagene. Buffers and other assay
reagents were purchased from either Sigma-Aldrich or Fisher Scientific
and were of the highest grade commercially available. Sequencing was
conducted by the University of Michigan DNA Sequencing Core Facility.

Andrews F.H., Rogers M.P., Paul L.N, & McLeish M.J. (2014). Perturbation of the Monomer–Monomer Interfaces of the Benzoylformate Decarboxylase Tetramer. Biochemistry, 53(27), 4358-4367.

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Purification of Recombinant LukAB Toxin

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A construct to co-purify recombinant LukAB from S. aureus (pOS-1-P_lukAB-sslukA-6His-lukA-lukB) was generated as previously described [24 (link)] and transformed into Newman ΔΔΔΔ [61 (link)] to facilitate purification. This construct was also used to individually express toxin subunits from LukED, PVL, HlgABC [24 (link)]. Toxins were purified from S. aureus as previously described [24 (link)]. Briefly, strains were grown in TSB with 10 μg/ml chloramphenicol for 5 h at 37°C, 180 rpm, to an optical density at 600 nm (OD₆₀₀) of approximately 1.5 (which represents 1 x 10⁹ CFU/ml). The bacteria were then pelleted, and the supernatant was collected and filtered. Nickel-nitrilotriacetic acid (NTA) resin (Qiagen) was incubated with culture supernatant, washed, and eluted with 500 mM imidazole. The protein was dialyzed in 1 × Tris-buffered saline (TBS) plus 10% glycerol at 4°C overnight and then stored at −80°C.

Melehani J.H., James D.B., DuMont A.L., Torres V.J, & Duncan J.A. (2015). Staphylococcus aureus Leukocidin A/B (LukAB) Kills Human Monocytes via Host NLRP3 and ASC when Extracellular, but Not Intracellular. PLoS Pathogens, 11(6), e1004970.

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