Histrap nickel column

Manufactured by GE Healthcare

Sourced in Germany

The HisTrap nickel column is a laboratory product designed for the purification of histidine-tagged proteins. It features a nickel-charged agarose matrix that selectively binds to the histidine tag, allowing the target protein to be isolated from a complex mixture. The column can be used in various protein purification workflows.

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Lab products found in correlation

Freestyle 293 expression medium, by Thermo Fisher Scientific (5 mentions) Expi293f cells, by Thermo Fisher Scientific (5 mentions) Superdex 75 10 300 gl gel filtration column, by GE Healthcare (5 mentions) Hek293t, by American Type Culture Collection (4 mentions) Strep tactin xt resin, by IBA Lifesciences (4 mentions) Hiload 16 60 superdex 75, by GE Healthcare (3 mentions) Edta free protease inhibitor tablet, by Roche (3 mentions) Superdex 75 hiload 16 60 gel filtration column, by GE Healthcare (2 mentions) Superdex 75 hiload 16 60 gel, by GE Healthcare (2 mentions) Superdex 200 size exclusion chromatography, by GE Healthcare (2 mentions) Kta pure, by GE Healthcare (2 mentions) D biotin, by Merck Group (2 mentions) Pdisplay bira er, by Addgene (2 mentions) Akta fplc, by GE Healthcare (2 mentions) Mlbugbuster protein extraction reagent, by Merck Group (2 mentions) Amicon ultra centrifugal filter, by Merck Group (1 mentions) Roller bottles, by Greiner (1 mentions) Hiprep, by Cytiva (1 mentions) Superdex 75 column, by GE Healthcare (1 mentions) Cation exchange chromatography, by GE Healthcare (1 mentions)

Spelling variants of this product

HisTrap column (331 citations) HisTrap (120 citations) Nickel column (27 citations) HisTrap HP nickel column (11 citations) HisTrap Nickel-affinity column (7 citations) HisTrap FF Nickel affinity column (3 citations) HisTrap FF nickel columns (2 citations) HisTrap excel nickel-affinity chromatography columns (2 citations) HisTrap FFTM nickel column (2 citations) HisTrap Sepharose nickel column (2 citations)

31 protocols using histrap nickel column

Omicron RBD and Spike Purification

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Protein expression and purification were conducted as described previously (Dejnirattisai et al., 2021a (link); Zhou et al., 2020 ). Briefly, plasmids encoding proteins were transiently expressed in HEK293T (ATCC CRL-11268) cells. The conditioned medium was concentrated using a QuixStand benchtop system. His-tagged Omicron RBD were purified with a 5 mL HisTrap nickel column (GE Healthcare) and further polished using a Superdex 75 HiLoad 16/60 gel filtration column (GE Healthcare). Twin-strep tagged Omicron spike was purified with Strep-Tactin XT resin (IBA lifesciences).
~4mg of ACE2 was mixed with homemade His-tagged 3C protease and DTT (final concentration 1mM). After incubated at 4 °C for one day, the sample was flown through a 5 mL HisTrap nickel column (GE Healthcare). His-tagged proteins were removed by the nickel column and purified ACE2 was harvested and concentrated.

Dejnirattisai W., Huo J., Zhou D., Zahradník J., Supasa P., Liu C., Duyvesteyn H.M., Ginn H.M., Mentzer A.J., Tuekprakhon A., Nutalai R., Wang B., Dijokaite A., Khan S., Avinoam O., Bahar M., Skelly D., Adele S., Johnson S.A., Amini A., Ritter T., Mason C., Dold C., Pan D., Assadi S., Bellass A., Omo-Dare N., Koeckerling D., Flaxman A., Jenkin D., Aley P.K., Voysey M., Clemens S.A., Naveca F.G., Nascimento V., Nascimento F., Fernandes da Costa C., Resende P.C., Pauvolid-Correa A., Siqueira M.M., Baillie V., Serafin N., Ditse Z., Da Silva K., Madhi S., Nunes M.C., Malik T., Openshaw P.J., Baillie J.K., Semple M.G., Townsend A.R., Huang K.Y., Tan T.K., Carroll M.W., Klenerman P., Barnes E., Dunachie S.J., Constantinides B., Webster H., Crook D., Pollard A.J., Lambe T., Paterson N.G., Williams M.A., Hall D.R., Fry E.E., Mongkolsapaya J., Ren J., Schreiber G., Stuart D.I, & Screaton G.R. (2021). Omicron-B.1.1.529 leads to widespread escape from neutralizing antibody responses. bioRxiv.

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Omicron Spike and ACE2 Purification

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Protein expression and purification were conducted as described previously (Dejnirattisai et al., 2021a (link); Zhou et al., 2020 (link)). Briefly, plasmids encoding proteins were transiently expressed in HEK293T (ATCC CRL-11268) cells. The conditioned medium was concentrated using a QuixStand benchtop system. His-tagged Omicron RBD were purified with a 5 mL HisTrap nickel column (GE Healthcare) and further polished using a Superdex 75 HiLoad 16/60 gel filtration column (GE Healthcare). Twin-strep tagged Omicron spike was purified with Strep-Tactin XT resin (IBA lifesciences).
∼4mg of ACE2 was mixed with homemade His-tagged 3C protease and DTT (final concentration 1mM). After incubation at 4 °C for one day, the sample was flowed through a 5 mL HisTrap nickel column (GE Healthcare). His-tagged proteins were removed by the nickel column and purified ACE2 was harvested and concentrated.

Dejnirattisai W., Huo J., Zhou D., Zahradník J., Supasa P., Liu C., Duyvesteyn H.M., Ginn H.M., Mentzer A.J., Tuekprakhon A., Nutalai R., Wang B., Dijokaite A., Khan S., Avinoam O., Bahar M., Skelly D., Adele S., Johnson S.A., Amini A., Ritter T.G., Mason C., Dold C., Pan D., Assadi S., Bellass A., Omo-Dare N., Koeckerling D., Flaxman A., Jenkin D., Aley P.K., Voysey M., Costa Clemens S.A., Naveca F.G., Nascimento V., Nascimento F., Fernandes da Costa C., Resende P.C., Pauvolid-Correa A., Siqueira M.M., Baillie V., Serafin N., Kwatra G., Da Silva K., Madhi S.A., Nunes M.C., Malik T., Openshaw P.J., Baillie J.K., Semple M.G., Townsend A.R., Huang K.Y., Tan T.K., Carroll M.W., Klenerman P., Barnes E., Dunachie S.J., Constantinides B., Webster H., Crook D., Pollard A.J., Lambe T., Paterson N.G., Williams M.A., Hall D.R., Fry E.E., Mongkolsapaya J., Ren J., Schreiber G., Stuart D.I, & Screaton G.R. (2022). SARS-CoV-2 Omicron-B.1.1.529 leads to widespread escape from neutralizing antibody responses. Cell, 185(3), 467-484.e15.

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Protein Expression and Purification

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Liu C., Zhou D., Nutalai R., Duyvesteyn H.M., Tuekprakhon A., Ginn H.M., Dejnirattisai W., Supasa P., Mentzer A.J., Wang B., Case J.B., Zhao Y., Skelly D.T., Chen R.E., Johnson S.A., Ritter T.G., Mason C., Malik T., Temperton N., Paterson N.G., Williams M.A., Hall D.R., Clare D.K., Howe A., Goulder P.J., Fry E.E., Diamond M.S., Mongkolsapaya J., Ren J., Stuart D.I, & Screaton G.R. (2022). The antibody response to SARS-CoV-2 Beta underscores the antigenic distance to other variants. Cell Host & Microbe, 30(1), 53-68.e12.

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Purification of GAAPs and hBI-1 in Yeast

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GAAPs and hBI-1 were expressed in S. cerevisiae strain FGY217 (34 (link)). The engineered proteins had a cleavable C-terminal GFP-His₈ tag and were expressed from the p424GAL1-TEVp-GFP-His₈ vector under control of the galactose promoter (35 (link)). The proteins were purified and analyzed according to a protocol developed for other transmembrane proteins (36 (link)) in 150 mm NaCl, 20 mm Tris-base, 5% glycerol, and 0.06% lauryldimethylamine N-oxide, pH 7.5. The GFP-His₈ tag was cleaved by adding His₈-tagged tobacco etch virus protease to the purified GFP-His₈-tagged protein at a molar ratio of 1:1 and digested overnight at 4 °C. Cleaved GFP-His₈ and the His-tagged protease were removed using a HisTrap nickel column (GE Healthcare), and the untagged target proteins were harvested from the flow-through. The purified proteins were concentrated using an Amicon Ultra centrifugal filter with a molecular mass cut-off of 30 kDa (Millipore) and analyzed on a Superdex 200 size exclusion chromatography (SEC) column (GE Healthcare). Fractions corresponding to the purified target protein were collected and concentrated to 1.5–2 mg/ml. Purified adenosine A_2A receptors (A_2ARs) were provided by Dr. S. Singh (37 (link)).

Carrara G., Saraiva N., Parsons M., Byrne B., Prole D.L., Taylor C.W, & Smith G.L. (2015). Golgi Anti-apoptotic Proteins Are Highly Conserved Ion Channels That Affect Apoptosis and Cell Migration. The Journal of Biological Chemistry, 290(18), 11785-11801.

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Expression and Purification of RBD Protein

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Stable HEK293S cell line expressing His-tagged RBD was cultured in DMEM (high glucose, Sigma) supplemented with 10% FBS (Invitrogen), 1 mM glutamine and 1x non-essential amino acids at 37 °C. Cells were transferred to roller bottles (Greiner) and cultured in DMEM supplemented with 2% FBS, 1 mM glutamine and 1x non-essential amino acids at 30 °C for 10 days for protein expression. For protein purification, the dialyzed media was passed through a 5 mL HisTrap Nickel column (GE Healthcare). The column was washed with buffer 20 mM Tris pH 7.4, 200 mM NaCl, 30 mM imidazole and RBD was eluted using buffer 20 mM Tris pH 7.4, 200 mM NaCl, 300 mM imidazole. A volume of 30 μL endoglycosidase H1 (~1 mg ml⁻¹) was added to ~30 mg RBD and incubated at room temperature for 2 h. Then the sample was further purified with a Superdex 75 HiLoad 16/600 gel filtration column (GE Healthcare) using 10 mM HEPES pH 7.4, 150 mM NaCl. Purified RBD was concentrated using a 10-kDa ultra centrifugal filter (Amicon) to 10.6 mg ml⁻¹ and stored at −80°C.

Dejnirattisai W., Zhou D., Ginn H.M., Duyvesteyn H.M., Supasa P., Case J.B., Zhao Y., Walter T.S., Mentzer A.J., Liu C., Wang B., Paesen G.C., Slon-Campos J., López-Camacho C., Kafai N.M., Bailey A.L., Chen R.E., Ying B., Thompson C., Bolton J., Fyfe A., Gupta S., Tan T.K., Gilbert-Jaramillo J., James W., Knight M., Carroll M.W., Skelly D., Dold C., Peng Y., Levin R., Dong T., Pollard A.J., Knight J.C., Klenerman P., Temperton N., Hall D.R., Williams M.A., Paterson N.G., Bertram F.K., Siebert C.A., Clare D.K., Howe A., Radecke J., Song Y., Townsend A.R., Huang K.Y., Fry E.E., Mongkolsapaya J., Diamond M.S., Ren J., Stuart D.I, & Screaton G.R. (2021). The antigenic anatomy of SARS-CoV-2 receptor binding domain. Cell, 184(8), 2183-2200.e22.

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Purification of Omicron and BA RBDs

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Protein expression and purification were conducted largely as described previously (Dejnirattisai et al., 2021a (link); Zhou et al., 2021 (link)). Twin-strep tagged Omicron spike was transiently expressed in HEK293T cells and purified with Strep-Tactin XT resin (IBA lifesciences). Plasmids encoding BA.1 RBD (319–541), BA.1 RBD (330–532) and BA.2 RBD (330–532) were transiently expressed in Expi293F™ Cells (ThermoFisher), cultured in FreeStyle™ 293 Expression Medium (ThermoFisher) at 30°C with 8% CO2 for 4 days. BA.1 RBD (330–532) was expressed in the presence of 1 μg/mL kifunensine. The harvested medium was concentrated using a QuixStand benchtop system. His-tagged ACE2 and RBDs were purified with a 5 mL HisTrap nickel column (GE Healthcare), followed by a Superdex 75 10/300 GL gel filtration column (GE Healthcare).

Nutalai R., Zhou D., Tuekprakhon A., Ginn H.M., Supasa P., Liu C., Huo J., Mentzer A.J., Duyvesteyn H.M., Dijokaite-Guraliuc A., Skelly D., Ritter T.G., Amini A., Bibi S., Adele S., Johnson S.A., Constantinides B., Webster H., Temperton N., Klenerman P., Barnes E., Dunachie S.J., Crook D., Pollard A.J., Lambe T., Goulder P., Paterson N.G., Williams M.A., Hall D.R., Mongkolsapaya J., Fry E.E., Dejnirattisai W., Ren J., Stuart D.I, & Screaton G.R. (2022). Potent cross-reactive antibodies following Omicron breakthrough in vaccinees. Cell, 185(12), 2116-2131.e18.

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Protein Production in HEK293T Cells

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Protein production was as described in Zhou et al. (2020 (link)). Briefly, plasmids encoding proteins were transiently expressed in HEK293T (ATCC CRL-11268) cells. The conditioned medium was dialysed and purified with a 5 mL HisTrap nickel column (GE Healthcare) and further polished using a Superdex 75 HiLoad 16/60 gel filtration column (GE Healthcare).

Dejnirattisai W., Zhou D., Supasa P., Liu C., Mentzer A.J., Ginn H.M., Zhao Y., Duyvesteyn H.M., Tuekprakhon A., Nutalai R., Wang B., López-Camacho C., Slon-Campos J., Walter T.S., Skelly D., Costa Clemens S.A., Naveca F.G., Nascimento V., Nascimento F., Fernandes da Costa C., Resende P.C., Pauvolid-Correa A., Siqueira M.M., Dold C., Levin R., Dong T., Pollard A.J., Knight J.C., Crook D., Lambe T., Clutterbuck E., Bibi S., Flaxman A., Bittaye M., Belij-Rammerstorfer S., Gilbert S.C., Carroll M.W., Klenerman P., Barnes E., Dunachie S.J., Paterson N.G., Williams M.A., Hall D.R., Hulswit R.J., Bowden T.A., Fry E.E., Mongkolsapaya J., Ren J., Stuart D.I, & Screaton G.R. (2021). Antibody evasion by the P.1 strain of SARS-CoV-2. Cell, 184(11), 2939-2954.e9.

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Recombinant Expression and Purification of HexD

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The plasmid
containing the gene encoding human HexD in a pET30a vector, which
has been reported previously,^{9 (link)} was transformed
into Escherichia coli BL21(DE3) cells. Successful
transformants were cultured in Luria-Bertani broth supplemented with
50 μg/mL kanamycin at 37 °C until an optical density of
0.6 absorbance units was reached. Protein expression was induced with
0.5 mM isopropyl β-d-thiogalactoside at 15 °C
for 20 h. Cells were harvested and resuspended in 20 mM HEPES (pH
7.4), 150 mM NaCl, and 5 mM imidazole and incubated in the presence
of 1 mg/mL lysozyme, 0.02 mg/mL DNase, and an EDTA-free protease inhibitor
tablet (Roche) for 20 min at 4 °C. Cells were lysed by high-pressure
cell disruption (Constant Systems). Following clarification, the supernatant
was applied to a 5 mL HisTrap nickel column (GE Healthcare), pre-equilibrated
in the same buffer, and the protein was eluted from an imidazole gradient.
The elution was concentrated and applied to a HiPrep 26/10 desalting
column equilibrated in 20 mM HEPES (pH 7.4), 150 mM NaCl buffer to
remove the imidazole. The elution was subsequently concentrated and
applied to a S200 16/60 gel filtration column, pre-equilibrated in
the same buffer, and fractions judged to be pure were pooled for subsequent
kinetic studies.

Alteen M.G., Oehler V., Nemčovičová I., Wilson I.B., Vocadlo D.J, & Gloster T.M. (2016). Mechanism of Human Nucleocytoplasmic Hexosaminidase D. Biochemistry, 55(19), 2735-2747.

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Purification of Monomeric α-Synuclein

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To get the
monomeric version of α-synuclein, purified α-syn-TEV Recognition
Site-GST-6His protein was transferred to the TEV reaction buffer (50
mM Tris-HCl [pH 8.0], 0.5 mM EDTA, and 1 mM DTT). TEV protease reaction
was done by using 500 μg/mL α syn, 100 μg/mL homemade
TEV protease-GST-6His, 0.02% sodium azide, and 0.1 mM DTT at room
temperature overnight. The monomeric proteins were then purified by
using the HisTrap nickel column (GE Life Sciences 17524701) for FPLC
(ÄKTA start protein purification system) as per the manufacturers’
specified protocol. The unbound proteins were collected since α-synuclein
had no tag to be captured by the HisTag column except TEV Protease-GST-6His
and GST-6His from α-syn-TEV Recognition Site-GST-6His. The monomeric
α-synuclein was transferred to 1× PBS and phosphate buffer
for QCM-D analysis and fibrillization assay, respectively.

Özçelik C.E., Beğli Ö., Hınçer A., Ahan R.E., Kesici M.S., Oğuz O., Kasırga T.S., Özçubukçu S, & Şeker U.Ö. (2023). Synergistic Screening of Peptide-Based Biotechnological Drug Candidates for Neurodegenerative Diseases Using Yeast Display and Phage Display. ACS Chemical Neuroscience, 14(19), 3609-3621.

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Structural Characterization of eOD-GT8 and Fab Complex

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For crystallography, a minimally glycosylated eOD-GT8 [His6x-tagged eOD-GT8minglyc (16 (link))] was produced by transiently transfected 293S (GnTI^−/−) cells and then purified using a HisTrap nickel column (GE Healthcare), followed by size exclusion chromatography using a Superdex75 column (GE Healthcare). The Fab constructs of the naive Abs were first affinity purified with human lambda resin (Life Technologies), followed by cation exchange chromatography (GE Healthcare). After incubating the purified Fab with a molar excess of purified eOD-GT8minglyc, the complex was deglycosylated by an Endo H treatment (New England Biolabs), followed by purification over Superdex200 size exclusion chromatography (GE Healthcare). The deglycosylated complex entered crystallization trials using an automated CrystalMation robotic system (Rigaku) at concentrations between 5 and 15 mg/ml. Crystals of the complex of the IOMA-class naive B cell CLK31 Fab with eOD-GT8minglyc (PDB ID: 6D2P; 2.60 Å resolution) were obtained at 20°C in a solution at pH 5.0 of 20% w/v PEG6000, and 0.1 M citric acid (pH 4.0). Crystals were cryoprotected in 25% glycerol followed by fast plunging in liquid nitrogen.

Havenar-Daughton C., Sarkar A., Kulp D.W., Toy L., Hu X., Deresa I., Kalyuzhniy O., Kaushik K., Upadhyay A.A., Menis S., Landais E., Cao L., Diedrich J.K., Kumar S., Schiffner T., Reiss S.M., Seumois G., Yates J.R., Paulson J.C., Bosinger S.E., Wilson I.A., Schief W.R, & Crotty S. (2018). The human naive B cell repertoire contains distinct subclasses for a germline-targeting HIV-1 vaccine immunogen. Science translational medicine, 10(448), eaat0381.

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