Nickel affinity column

Manufactured by Thermo Fisher Scientific

Sourced in United States

Nickel affinity columns are a type of chromatography column used for the purification of His-tagged proteins. They contain a nickel-charged resin that binds to the histidine (His) tag on the target protein, allowing it to be separated from other proteins in a sample. The core function of nickel affinity columns is to facilitate the isolation and purification of His-tagged proteins.

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Lab products found in correlation

Freestyle 293f medium, by Thermo Fisher Scientific (13 mentions) Freestyle 293 f cells, by Thermo Fisher Scientific (13 mentions) Expifectamine 293 transfection reagent, by Thermo Fisher Scientific (12 mentions) 0.22 μm filter, by Thermo Fisher Scientific (10 mentions) Protein a affinity column, by Cytiva (2 mentions) 0.22 m filter, by Thermo Fisher Scientific (2 mentions) Anti his antibody, by Abmart (1 mentions) Restriction enzyme, by Takara Bio (1 mentions) Dna polymerase, by Thermo Fisher Scientific (1 mentions) Gly pro p nitroanilide, by Merck Group (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Akta prime plus fplc, by GE Healthcare (1 mentions) Hiload 16 60 superdex 200 pg, by GE Healthcare (1 mentions) Hek293t cell, by American Type Culture Collection (1 mentions) Ion exchange column, by GE Healthcare (1 mentions)

Close spelling variants of this product

Nickel column (2 citations)

16 protocols using nickel affinity column

SARS-CoV-2 Spike RBD Protein Production

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FreeStyle 293F cells (Invitrogen) were grown in FreeStyle 293F medium (Invitrogen) to a density of 1 × 10⁶ cells/mL at 37 °C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for SARS-CoV-2 S RBD [32 (link)] using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 μm filter (Thermo Fisher Scientific, Waltham, MA, USA). The recombinant RBD proteins were purified by nickel affinity columns, as directed by the manufacturer (Invitrogen). The RBD preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at −80 °C until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie Blue.

Marchitto L., Chatterjee D., Ding S., Gendron-Lepage G., Tauzin A., Boutin M., Benlarbi M., Medjahed H., Sylla M., Lanctôt H., Durand M., Finzi A, & Tremblay C. (2023). Humoral Responses Elicited by SARS-CoV-2 mRNA Vaccine in People Living with HIV. Viruses, 15(10), 2004.

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Production of SARS-CoV-2 Spike RBD Protein

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FreeStyle 293 F cells (Invitrogen) were grown in FreeStyle 293F medium (Invitrogen) to a density of 1 × 10⁶ cells/mL at 37 °C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for SARS-CoV-2 S RBD using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 μm filter (Thermo Fisher Scientific, Waltham, MA, USA). The recombinant RBD proteins were purified by nickel affinity columns, as directed by the manufacturer (Invitrogen). The RBD preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at −80 °C until further use. To assess purity, recombinant proteins were loaded on SDSPAGE gels and stained with Coomassie Blue.

Chatterjee D., Tauzin A., Laumaea A., Gong S.Y., Bo Y., Guilbault A., Goyette G., Bourassa C., Gendron-Lepage G., Medjahed H., Richard J., Moreira S., Côté M, & Finzi A. (2022). Antigenicity of the Mu (B.1.621) and A.2.5 SARS-CoV-2 Spikes. Viruses, 14(1), 144.

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Recombinant SARS-CoV-2 RBD Protein Production

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FreeStyle 293 F cells (Invitrogen) were grown in FreeStyle 293F medium (Invitrogen) to a density of 10⁶ cells/mL at 37 °C with 8% CO₂ under regular agitation (150 rpm). Cells were transfected with the plasmid coding for SARS-CoV-2 S RBD WT using an ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 μm filter (Thermo Fisher Scientific, Waltham, MA, USA). The recombinant RBD proteins were purified using nickel affinity columns, as directed by the manufacturer (Invitrogen). The RBD preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at −80 °C until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie Blue.

Tauzin A., Gendron-Lepage G., Nayrac M., Anand S.P., Bourassa C., Medjahed H., Goyette G., Dubé M., Bazin R., Kaufmann D.E, & Finzi A. (2022). Evolution of Anti-RBD IgG Avidity following SARS-CoV-2 Infection. Viruses, 14(3), 532.

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Recombinant ACE2 Protein Production

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FreeStyle 293F cells (Invitrogen, Rockford, IL, USA) were grown in FreeStyle 293F medium (Invitrogen) to a density of 1 × 10⁶ cells/mL at 37 °C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for soluble ACE2 or ACE2-Fc using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 µm filter (Thermo Fisher Scientific, Waltham, MA, USA). The recombinant sACE2 protein was purified by nickel affinity columns (Invitrogen) and ACE2-Fc was purified using protein A affinity column (Cytiva, Marlborough, MA, USA), as directed by the manufacturers. The protein preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at −80 °C until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie blue.

Anand S.P., Chen Y., Prévost J., Gasser R., Beaudoin-Bussières G., Abrams C.F., Pazgier M, & Finzi A. (2020). Interaction of Human ACE2 to Membrane-Bound SARS-CoV-1 and SARS-CoV-2 S Glycoproteins. Viruses, 12(10), 1104.

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Recombinant SARS-CoV-2 RBD Protein Production

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FreeStyle 293F cells (Invitrogen) were grown in FreeStyle 293F medium (Invitrogen) to a density of 1 × 10⁶ cells/mL at 37°C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for SARS-CoV-2 S WT RBD (Beaudoin-Bussières et al., 2020 (link)) using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 μm filter (Thermo Fisher Scientific). The recombinant RBD proteins were purified by nickel affinity columns, as directed by the manufacturer (Invitrogen). The RBD preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at −80°C until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie Blue.

Tauzin A., Gong S.Y., Chatterjee D., Ding S., Painter M.M., Goel R.R., Beaudoin-Bussières G., Marchitto L., Boutin M., Laumaea A., Okeny J., Gendron-Lepage G., Bourassa C., Medjahed H., Goyette G., Williams J.C., Bo Y., Gokool L., Morrisseau C., Arlotto P., Bazin R., Fafard J., Tremblay C., Kaufmann D.E., De Serres G., Richard J., Côté M., Duerr R., Martel-Laferrière V., Greenplate A.R., Wherry E.J, & Finzi A. (2022). A boost with SARS-CoV-2 BNT162b2 mRNA vaccine elicits strong humoral responses independently of the interval between the first two doses. Cell Reports, 41(4), 111554.

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DPP Enzymatic Activity Assay

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The recognised dipeptidyl peptidase (DPP) substrate Gly-Pro-p-nitroanilide and DPP4 enzyme were purchased from Sigma (Missouri, USA). The VigoScript first strand cDNA synthesis kit was a product of Vigorous (Beijing, China). Rosetta competent cells were obtained from TransGen (Beijing, China). Restriction enzymes were purchased from Takara (Dalian, China). Nickel affinity columns, Trizol reagent and DNA polymerase were obtained from Invitrogen (Shanghai, China). The plasmid vector pET32-a(+) was a product of Novagen (Shanghai, China). The anti-His antibody was purchased from Abmart (Shanghai, China). The DPP inhibitor compounds, sitagliptin and UAMC00132 were provided by Prof. Haihong Huang.

Liu J., Huan Y., Li C., Liu M, & Shen Z. (2014). Establishment of a selective evaluation method for DPP4 inhibitors based on recombinant human DPP8 and DPP9 proteins. Acta Pharmaceutica Sinica. B, 4(2), 135-140.

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Recombinant SARS-CoV-2 RBD Protein Production

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FreeStyle 293F cells (Invitrogen) were grown in FreeStyle 293F medium (Invitrogen) to a density of 1 × 10⁶ cells/mL at 37°C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for SARS-CoV-2 S RBD (Beaudoin-Bussières et al., 2020 (link)) using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 μm filter (Thermo Fisher Scientific). The recombinant RBD proteins were purified by nickel affinity columns, as directed by the manufacturer (Invitrogen). The RBD preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at −80°C until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie Blue.

Tauzin A., Beaudoin-Bussières G., Gong S.Y., Chatterjee D., Gendron-Lepage G., Bourassa C., Goyette G., Racine N., Khrifi Z., Turgeon J., Tremblay C., Martel-Laferrière V., Kaufmann D.E., Cardinal H., Cloutier M., Bazin R., Duerr R., Dieudé M., Hébert M.J, & Finzi A. (2022). Humoral immune responses against SARS-CoV-2 Spike variants after mRNA vaccination in solid organ transplant recipients. iScience, 25(9), 104990.

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SARS-CoV-2 RBD Protein Production

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FreeStyle 293F cells (Thermo Fisher Scientific) were grown in FreeStyle 293F medium (Thermo Fisher Scientific) to a density of 1 × 10⁶ cells/mL at 37°C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for SARS-CoV-2 S RBD using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Thermo Fisher Scientific). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 μm filter (Thermo Fisher Scientific). The recombinant RBD proteins were purified by nickel affinity columns, as directed by the manufacturer (Invitrogen). The RBD preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at −80°C until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie Blue.

Anand S.P., Prévost J., Nayrac M., Beaudoin-Bussières G., Benlarbi M., Gasser R., Brassard N., Laumaea A., Gong S.Y., Bourassa C., Brunet-Ratnasingham E., Medjahed H., Gendron-Lepage G., Goyette G., Gokool L., Morrisseau C., Bégin P., Martel-Laferrière V., Tremblay C., Richard J., Bazin R., Duerr R., Kaufmann D.E, & Finzi A. (2021). Longitudinal analysis of humoral immunity against SARS-CoV-2 Spike in convalescent individuals up to 8 months post-symptom onset. Cell Reports Medicine, 2(6), 100290.

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Recombinant SARS-CoV-2 Omicron BA.2 Spike Protein Production

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FreeStyle 293F cells (Invitrogen, Waltham, MA, USA) were grown in FreeStyle 293F medium (Invitrogen) to a density of 1 × 10⁶ cells/mL at 37 °C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for SARS-CoV-2 Omicron BA.2 S RBD (319-537), soluble ACE2 (sACE2, 1-615), or ACE2-Fc (1-615), using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 µm filter (Thermo Fisher Scientific, Waltham, MA, USA). The recombinant sACE2 protein and RBD proteins were purified by nickel affinity columns, as directed by the manufacturer (Invitrogen) and ACE2-Fc was purified using a protein A affinity column (Cytiva, Marlborough, MA, USA), as directed by the manufacturer. The protein preparations were dialysed against phosphate-buffered saline (PBS) and stored at −80 °C in aliquots until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie Blue. Purified proteins were >95% pure after size-exclusion chromatography as verified by SDS-PAGE and Coomassie blue staining.

Gong S.Y., Ding S., Benlarbi M., Chen Y., Vézina D., Marchitto L., Beaudoin-Bussières G., Goyette G., Bourassa C., Bo Y., Medjahed H., Levade I., Pazgier M., Côté M., Richard J., Prévost J, & Finzi A. (2022). Temperature Influences the Interaction between SARS-CoV-2 Spike from Omicron Subvariants and Human ACE2. Viruses, 14(10), 2178.

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Recombinant SARS-CoV-2 RBD Protein Production

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FreeStyle 293 F cells (Invitrogen) were grown in FreeStyle 293F medium (Invitrogen) to a density of 1 × 10⁶ cells/mL at 37°C with 8% CO₂ with regular agitation (150 rpm). Cells were transfected with a plasmid coding for SARS-CoV-2 S RBD using ExpiFectamine 293 transfection reagent, as directed by the manufacturer (Invitrogen). One week later, cells were pelleted and discarded. Supernatants were filtered using a 0.22 μm filter (Thermo Fisher Scientific). The recombinant RBD proteins were purified by nickel affinity columns, as directed by the manufacturer (Invitrogen). The RBD preparations were dialyzed against phosphate-buffered saline (PBS) and stored in aliquots at -80°C until further use. To assess purity, recombinant proteins were loaded on SDS-PAGE gels and stained with Coomassie Blue.

Gong S.Y., Chatterjee D., Richard J., Prévost J., Tauzin A., Gasser R., Bo Y., Vézina D., Goyette G., Gendron-Lepage G., Medjahed H., Roger M., Côté M, & Finzi A. (2021). Contribution of single mutations to selected SARS-CoV-2 emerging variants spike antigenicity. Virology, 563, 134-145.

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