Histrap column

Manufactured by GE Healthcare

Sourced in United States, United Kingdom, Sweden, France

The HisTrap column is a versatile affinity chromatography tool designed for the purification of histidine-tagged recombinant proteins. It features a prepacked matrix that selectively binds to the histidine-tag, allowing for efficient capture and purification of the target protein.

Automatically generated - may contain errors

Lab products found in correlation

Hitrap, by Cytiva (26 mentions) Pd 10 column, by GE Healthcare (7 mentions) Hitrap desalting column, by GE Healthcare (7 mentions) Superdex 200 column, by GE Healthcare (7 mentions) Hitrap q column, by GE Healthcare (6 mentions) Superdex 75 column, by GE Healthcare (6 mentions) Superdex 75, by GE Healthcare (5 mentions) Superdex 200 16 60 column, by GE Healthcare (5 mentions) Amicon ultra centrifugal filter, by Merck Group (5 mentions) Heparin column, by GE Healthcare (5 mentions) Superdex 75 10 300 column, by GE Healthcare (4 mentions) Freestyle 293 f, by Thermo Fisher Scientific (4 mentions) Pd 10 desalting column, by GE Healthcare (4 mentions) Hitrap q hp column, by GE Healthcare (4 mentions) Protease inhibitor cocktail, by Roche (4 mentions) Superdex 200, by GE Healthcare (4 mentions) Superdex 200 10 300 gl column, by GE Healthcare (4 mentions) Hiload 26 600 superdex200 pg, by GE Healthcare (4 mentions) Size exclusion columns, by GE Healthcare (4 mentions) Hitrap sp column, by GE Healthcare (3 mentions)

Spelling variants of this product

HisTrap (120 citations) HisTrap Fast Flow column (13 citations) HisTrap™ High Performance 1 mL columns (3 citations) HisTrap HP histidine-tagged protein columns (2 citations) HisTrap™ HP Crude Columns (2 citations) HisTrap excel nickel-affinity chromatography columns (2 citations) HisTrap TALON FF crude columns (2 citations) HisTrap HP Ni2+ Sepharose columns (2 citations) HisTrap FF nickel columns (2 citations) HisTrap™ FF Ni Sepharose Columns (2 citations)

331 protocols using histrap column

Recombinant Expression and Purification of Krimper eTud Domains

Check if the same lab product or an alternative is used in the 5 most similar protocols

Krimper eTud1 (residues 272-512) was cloned into a self-modified pSumo vector with 10×His tag followed by a yeast sumo sequence. The plasmid was transformed into E. coli strain BL21(DE3) Rosseta and cultured at 37 °C in LB medium. The protein expression was induced by adding IPTG to a final concentration of 0.2 mM when the OD₆₀₀ reached 0.7, and the cells were cooled to 16 °C. The recombinant expressed protein was purified using a HisTrap column (GE Healthcare). The hexahistidine plus yeast sumo tag was removed by ulp1 protease digestion followed by a second step HisTrap column (GE Healthcare). The target protein was further purified using MonoQ and Superdex G75 columns (GE Healthcare).
Krimper eTud2 (residues 562-746) was cloned into a self-modified His-MBP vector. Protein production procedure was to with Krimper eTud1. The recombinant expressed protein was purified using a HisTrap column (GE Healthcare). The hexahistidine plus MBP tag was removed by TEV protease digestion followed by a second step HisTrap column (GE Healthcare). Protein was further purified using Q and Superdex G75 columns (GE Healthcare).

Huang X., Hu H., Webster A., Zou F., Du J., Patel D.J., Sachidanandam R., Toth K.F., Aravin A.A, & Li S. (2021). Binding of guide piRNA triggers methylation of the unstructured N-terminal region of Aub leading to assembly of the piRNA amplification complex. Nature Communications, 12, 4061.

+ Open protocol

+ Expand

Expression and Purification of mTC/S Proteins

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Recombinant native and variant mTC/S proteins were expressed in E. coli Arctic Express (DE3). Cells were freshly transformed with a pET-mTC/S plasmid, grown in LB medium and induced with 0.1 mM IPTG for 16 h at 12° C according to the manufacturer’s instructions. The recombinant proteins were purified as described previously13 (link). Briefly, the cells were harvested and resuspended in buffer A (25 mM Tris pH 8.0, 150 mM NaCl, 1 mM DTT, 4 mM MgCl₂ and 5% (v/v) glycerol). The cells were lysed by sonication and the debris was removed by centrifugation. The supernatant was loaded onto a His-Trap column (GE Healthcare) pre-equilibrated with buffer A containing 25 mM imidazole. The column was washed with buffer A containing 25 mM imidazole, and the proteins were eluted by increasing the imidazole concentration to 500 mM. The purified proteins were desalted prior to removal of the His-tag by overnight incubation with TEV protease at 4°C with gentle mixing. The TEV protease was removed by passing the protein mixtures through a His-Trap column, and the proteins collected in the flow-through were concentrated and loaded onto a Hiload Superdex (26/60) S200 column (GE Healthcare) pre-equilibrated with buffer A. Purified protein fractions were concentrated to 8-10 mg/ml, aliquoted and stored at -80°C.

Leferink N.G., Ranaghan K.E., Karuppiah V., Currin A., van der Kamp M.W., Mulholland A.J, & Scrutton N.S. (2018). Experiment and Simulation Reveal How Mutations in Functional Plasticity Regions Guide Plant Monoterpene Synthase Product Outcome. ACS catalysis, 8(5), 3780-3791.

+ Open protocol

+ Expand

Seleno-methionine Protein Expression Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

To ensure efficient incorporation of seleno-methionine into the expressed protein, B834 (DE3) cells (Novagen) were used. An overnight culture was grown at 37°C in LB. Unlabelled media (Molecular Dimesions) was inoculated with 10ml of overnight culture and grown until OD₆₀₀ reached 0.5. Cells were harvested and washed three times in PBS before resuspending the pellets in seleno-methionine labelled media (Molecular Dimension). After a 40 minute incubation at 20°C, cells were induced with 0.4mM IPTG and left to express for 16 hours.
Harvested cells were lysed using a microfluidizer at 20,000 psi in a buffer containing 50mM Tris pH 8.0, 300mM NaCl, and 5mM BME. Cell lysates were clarified by centrifugation at 20,000xg and filtered through a 0.45μm filter. Lysates were then loaded onto a HisTrap column (GE Healthcare) before washing with 20mM imidazole. The protein was eluted with a linear gradient of imidazole from 0-300mM. Fractions containing protein were pooled and dialysed overnight at 4°C in the presence of TEV in a buffer containing 20mM Tris pH 8.0, 150mM NaCl, 5mM imidazole, and 2mM BME. The following day, proteins were passed over a HisTrap column and the flowthrough collected which was concentrated and passed over a Superdex S200 (GE Healthcare) column equilibrated in 10mM Tris pH7.5, 50mM NaCl and 2mM DTT.

Yelland T., Le A.H., Nikolaou S., Insall R., Machesky L, & Ismail S. (2021). Structural Basis of CYRI-B Direct Competition with Scar/WAVE Complex for Rac1. Structure(London, England:1993), 29(3), 226-237.e4.

+ Open protocol

+ Expand

Purification of SpaD Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

The SpaD proteins were purified using similar procedures to those used for the purification of SpaA (Kang, Paterson & Baker, 2009 ▶ ). Briefly, cleared cell lysate was loaded onto a 5 ml HisTrap column (GE Healthcare) charged with Ni²⁺, and the His-tagged SpaD proteins were eluted with a gradient of 20–500 mM imidazole. To remove the His tag, the eluted proteins were mixed with human rhinovirus (HRV) 3C protease (1 mg per ∼50 mg SpaD), 10 mM DTT and 2 mM EDTA and dialyzed against 50 mM Tris–HCl pH 8.0, 300 mM NaCl overnight at 277 K. For SeMet-substituted SpaD, the buffers were supplemented with 0.4 mM DTT from this step to prevent selenomethionine oxidation. The dialyzed sample was passed through a charged HisTrap column to collect the flowthrough containing untagged SpaD. Further purification was carried out using size-exclusion chromatography (SEC) on a Superdex 75 10/300 column (GE Healthcare) with SEC buffer (10 mM Tris–HCl pH 8.0, 50 mM NaCl). The final product contained two additional N-terminal residues, Gly and Pro, after His-tag removal.

Kang H.J., Paterson N.G., Kim C.U., Middleditch M., Chang C., Ton-That H, & Baker E.N. (2014). A slow-forming isopeptide bond in the structure of the major pilin SpaD from Corynebacterium diphtheriae has implications for pilus assembly. Acta Crystallographica Section D: Biological Crystallography, 70(Pt 5), 1190-1201.

+ Open protocol

+ Expand

Purification of DnaK and WT::NR Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

To purify the WT and point-mutant DnaK proteins, cells were pelleted after induction by centrifugation at 5,000 g and resuspended in ice-cold 2X-PBS (20 mM Na₂HPO4, 1.76mM KH₂PO4, 274mM NaCl and 5.4mM KCl). All purification steps were performed at 4°C. Before loading onto a HisTrap column (GE Healthcare), sonication was done in ice-water bath for 3 minutes followed by centrifugation at 12,000 g. DnaK proteins eluted from the HisTrap column were first dialyzed again 2X-PBS overnight, further purified with a HiTrap Q column (GE Healthcare), and evaluated by PAGE (Fig. S1A). Fractions containing DnaK protein were pooled, concentrated to >20mg/ml in a buffer containing 10 mM Hepes-KOH, pH 7.5, and 50 mM KCl, and flash frozen in liquid nitrogen.
The WT∷NR fusion protein was first purified using a HisTrap column as described above. After dialysis against 2X-PBS overnight, the Smt3 tag was cleaved off by Ulp1 protease, and removed by loading onto a HisTrap column. The WT∷NR fusion protein was further purified with a HiTrap Q column as described above, and a Superdex 200 column (26/60), saving the monomeric fraction (Fig. S1B). After concentration to >20mg/ml, samples were flash frozen in liquid nitrogen.

Wang W., Liu Q., Liu Q, & Hendrickson W.A. (2021). Conformational Equilibria in Allosteric Control of Hsp70 Chaperones. Molecular cell, 81(19), 3919-3933.e7.

+ Open protocol

+ Expand

Purification of NBX Fusion Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 17

TEV protease-cleavable, 6×His-thioredoxin-NBX fusion proteins are expressed in the cytoplasm of E. coli grown in autoinducing media (Formedium) for 24 hours at 30° C. Bacteria are collected by centrifugation, resuspended in buffer A (10 mM HEPES, pH 7.5, 500 mM NaCl, 20 mM Imidazole) and lysed using homogenization. Insoluble material is removed by centrifugation and the remaining soluble fraction is applied to a HisTrap column (GE Biosciences) pre-equilibrated with buffer A. The protein is eluted from the column using an FPLC with a linear gradient between buffer A and buffer B (10 mM HEPES, pH 7.5, 500 mM NaCl, 500 mM Imidazole). The eluted protein is dialyzed overnight in the presence of TEV protease to buffer C (10 mM HEPES, pH 7.5, 500 mM NaCl). The dialyzed protein is applied to a HisTrap column (GE Biosciences) pre-equilibrated with buffer C. 6×His-tagged TEV and 6×His-tagged thioredoxin are bound to the column and highly purified NBX is collected in the flowthrough. NBX proteins are dialyzed overnight to PBS and concentrated to −10 mg/ml.

US11130800B2. Antibodies against microorganisms and uses thereof (2021-09-28). NOVOBIND LIVESTOCK THERAPEUTICS INC. [CA]. Inventors: Hamlet Abnousi [CA].

+ Open protocol

+ Expand

Expression and Purification of Active S6K1

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human RPS6KB1 gene fragment, encoding residues 75–399, was PCR-amplified and cloned into the pDEST 10 vector (Invitrogen). The plasmid thus generated was transformed into DH10 Bac competent cells, and the recombinant bacmid was extracted and transfected into Sf9 insect cells. The Sf9 cells were infected with the baculovirus at a multiplicity of infection of one, and were incubated at 27 °C for 48 h. The Sf9 cells were collected, frozen, and stored at −80 °C until use. The cells were lysed using a sonicator, and the supernatant was applied to a HisTrap column (GE Healthcare). The target protein was eluted and treated with λ protein phosphatase, to remove non-specific phosphorylation, and the N-terminal 6× His tag was removed by TEV digestion. The protein was reloaded on the HisTrap column to remove the undigested protein, and the flow-through fractions were further purified by chromatography on a HiTrap SP column (GE Healthcare) and a HiLoad 16/60 Superdex 75 column (GE Healthcare).
Phosphorylation of the purified S6K1KD protein, using PDK1, was performed as previously described [17 (link)]. The His-tagged PDK1 protein was produced using a baculovirus expression system, and was purified prior to addition to S6K1KD.

Niwa H., Mikuni J., Sasaki S., Tomabechi Y., Honda K., Ikeda M., Ohsawa N., Wakiyama M., Handa N., Shirouzu M., Honma T., Tanaka A, & Yokoyama S. (2014). Crystal structures of the S6K1 kinase domain in complexes with inhibitors. Journal of Structural and Functional Genomics, 15(3), 153-164.

+ Open protocol

+ Expand

Purification of Soluble DSG3 Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Soluble mouse DSG3-His proteins were expressed in DG44 and affinity purified from the supernatants of the transfectants with His-Trap column (GE Healthcare).
Soluble human DSG3 was expressed as sDSG3-mIgG2aFc composed of the extracellular domain of human DSG3 (aa 1-616) and the Fc portion of mouse IgG2a, and was purified with HiTrap Protein G HP column (GE Healthcare) and eluted with 0.1 M Glycine-HCl (pH 2.7). The eluate was gel-filtrated with Superdex 200HR 10/30 (GE Healthcare).
Glutathione S-transferase (GST)-human DSG3 fusion protein (GST-hDSG3) was expressed in Escherichia coli as a fusion of GST and 125 aa of human DSG3 (aa 491-615) with His-tag. GST-hDSG3 was purified with His-Trap column (GE Healthcare) for use as an antigen for ELISA.

Funahashi S.I., Kawai S., Fujii E., Taniguchi K., Nakano K., Ishikawa S., Aburatani H, & Suzuki M. (2018). Generation of an anti-desmoglein 3 antibody without pathogenic activity of pemphigus vulgaris for therapeutic application to squamous cell carcinoma. Journal of Biochemistry, 164(6), 471-481.

+ Open protocol

+ Expand

Expression and Purification of Arabidopsis JMJ13 Catalytic Domain

Check if the same lab product or an alternative is used in the 5 most similar protocols

The catalytic fragment of Arabidopsis JMJ13 (JMJ13CD, residues 90–578) was cloned into a pET-Sumo vector to fuse an N-terminal hexahistidine plus yeast sumo tag. The plasmid was transformed into E. coli strain BL21(DE3) RIL and the transformants were cultured at 37 °C in LB medium. When the OD₆₀₀ of cell culture reached 0.7, the protein expression was induced by adding IPTG to a final concentration of 0.2 mM and the cells were cooled to 20 °C. The recombinant expressed protein was purified using a HisTrap column (GE Healthcare). The hexahistidine plus yeast sumo tag was removed by ulp1 protease digestion followed by a second step HisTrap column (GE Healthcare). The target protein was further purified on Heparin and Superdex G200 columns (GE Healthcare). The untagged JMJ13CD easily precipitates in the in vitro activity assay. We further cloned JMJ13CD into a pMal vector (New England Biolabs) to fuse an MBP tag to the target protein. The MBP-tagged JMJ13CD was expressed in E. coli strain BL21(DE3) RIL with IPTG induction and purified using amylose resin (New England Biolabs), Heparin, and Superdex G200 columns (GE Healthcare). All the mutations were generated using a PCR based method and purified using the same protocol as wild-type protein. The chemicals and peptides were purchased from Sigma-Aldrich and GL Biochem Company, respectively.

Zheng S., Hu H., Ren H., Yang Z., Qiu Q., Qi W., Liu X., Chen X., Cui X., Li S., Zhou B., Sun D., Cao X, & Du J. (2019). The Arabidopsis H3K27me3 demethylase JUMONJI 13 is a temperature and photoperiod dependent flowering repressor. Nature Communications, 10, 1303.

+ Open protocol

+ Expand

Purification of RNAP holoenzyme and phage proteins

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The RNAP holoenzyme used for crystallization was purified from T. thermophilus HB8 cells, as described previously (14 (link)). The phage proteins gp39 and gp76 were expressed in Escherichia coli, and purified as described (12 (link)). For the preparation of the SeMet derivative of gp76, the single amino acid substitution of Met for Leu at position 13 (L13M) was performed, as gp76 only contains an N-terminal Met residue. To obtain the SeMet derivative of the L13M variant, E. coli cells bearing the expression vector were cultured in M9 medium containing SeMet (15 (link)). The His-tagged proteins were purified by chromatography on a HisTrap column (GE Healthcare), and then the N-terminal His-tags were removed by thrombin cleavage followed by another purification with the HisTrap column. The protein was further purified by Superdex75 gel filtration column chromatography (GE Healthcare).

Ooi W.Y., Murayama Y., Mekler V., Minakhin L., Severinov K., Yokoyama S, & Sekine S.I. (2017). A Thermus phage protein inhibits host RNA polymerase by preventing template DNA strand loading during open promoter complex formation. Nucleic Acids Research, 46(1), 431-441.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!