Hitrap q hp column

Manufactured by GE Healthcare

Sourced in United States, Sweden, United Kingdom

The HiTrap Q HP column is a small-scale ion exchange chromatography column designed for fast and efficient purification of biomolecules. It features a strong anion exchange (quaternary ammonium) resin that can bind and separate negatively charged molecules based on their charge and size.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

HiTrap SP/Q HP columns (3 citations) HiTrap Q HP (124 citations) HiTrap HP column (20 citations) HiTrap Q column (111 citations) HiTrap Q (51 citations) HiTrap columns (11 citations)

176 protocols using hitrap q hp column

Conjugation of pG to SMCC-modified ODN

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

For the conjugation of pG to a SMCC-functionalized
ODN, an aliquot
of pG was buffer-exchanged to (100 mM sodium phosphate, 25 μM
TCEP, pH 7) using a PD10 desalting column (GE Healthcare). Subsequently,
desalted pG was concentrated to a final concentration of 50 μM
using Amicon 3 kDa MWCO centrifugal filters (Merck Millipore). Ten
nmol lyophilized SMCC-functionalized ODN was reconstituted in 40 μL
of 50 μM pG (2 nmol), resulting in a five times excess of maleimide–ODN.
The reaction was shaken at 850 rpm for 3 h at 20 °C. The coupling
efficiency was assessed using SDS-PAGE under nonreducing conditions.
The purification, if applicable, of pG-ODN was performed using fast
protein liquid chromatography (FPLC, ÄKTA Prime, GE Healthcare)
with an anion-exchange HiTrap Q HP column (1 mL, GE Healthcare) using
a salt gradient with a start and end concentration of 100 and 500
mM NaCl in 50 mM Tris-HCl (pH 7.5), respectively. Elution fractions
were collected and analyzed by measuring online absorption at 280
nm and SDS-PAGE under nonreducing conditions. pG-ODN conjugates were
aliquoted and stored at −80 °C.

Cremers G.A., Rosier B.J., Riera Brillas R., Albertazzi L, & de Greef T.F. (2019). Efficient Small-Scale Conjugation of DNA to Primary Antibodies for Multiplexed Cellular Targeting. Bioconjugate Chemistry, 30(9), 2384-2392.

+ Open protocol

+ Expand

Cell Fractionation and Extraction

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

Hypotonic lysates and nuclear extracts were prepared using standard techniques described previously (Wang et al., 1994 (link)). Where noted, cell lysates were fractionated by binding to a 1 mL HiTrap Q HP column (GE Healthcare) in 20 mM bis-tris-propane-HCl pH 8.7, 5 mM KCl, 1 mM MgCl₂ and eluting with increasing concentrations of KCl.

Hosios A.M., Fiske B.P., Gui D.Y, & Vander Heiden M.G. (2015). Lack of evidence for PKM2 protein kinase activity. Molecular cell, 59(5), 850-857.

+ Open protocol

+ Expand

Purification of UvrA Protein from Thermotoga maritima

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

UvrA gene from Thermotoga maritima was encoded into the pET28a (+) vector and transformed into E. coli Rosetta (DE3) pLysS cells. Protein expression was induced using 50 µM isopropyl-ß-d-galactoside (IPTG) and incubated for 5 h at 30 °C. Bacterial cells were resuspended in buffer containing 50 mM Tris pH 8.0, 150 mM NaCl, 2 mM dithiothreitol (DTT), 1 mM phenylmethylsulfonyl fluoride (PMSF) and 5% (v/v) glycerol and sonicated. The lysate was centrifuged at 18,000 rpm for 45 min and the supernatant was applied on His-Trap crude 5 ml column (GE Healthcare). UvrA was eluted with a 200 ml imidazole gradient from 0 to 500 mM concentration. The purified protein was diluted with buffer containing 50 mM Tris pH 8.0, 2 mM DTT, and 5% (v/v) glycerol and applied to 5 mL HiTrap Q HP column (GE Healthcare). UvrA was eluted with 50 ml linear NaCl gradient from 50 mM to 1 M. The protein was further purified by size exclusion chromatography using Superdex 200 10/300 GL (GE Healthcare) equilibrated with buffer containing 50 mM Tris pH 8.0, 200 mM NaCl, 2 mM DTT and 5% (v/v) glycerol. The volume of the collected protein sample was reduced to a final concentration of 10 mg/ml and flash-frozen in liquid nitrogen.

Gade P., Erlandson A., Ullah A., Chen X., Mathews I.I., Mera P.E, & Kim C.Y. (2023). Structural and functional analyses of the echinomycin resistance conferring protein Ecm16 from Streptomyces lasalocidi. Scientific Reports, 13, 7980.

+ Open protocol

+ Expand

Purification of ADAT2/ADAT3 Complex and TadA from E. coli

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

ADAT2/ADAT3 WT and mutants were produced by co-expression in Escherichia coli BL21(DE3) cells in LB Broth medium. Culture induction was performed at 22°C by adding final concentration of 0.5 mM of isopropyl-1-thio-β-D-galactopyranoside (IPTG) in presence of 100 μM of Zn(SO₄)₂. Cells were harvested, resuspended and lysed in a buffer containing 10 mM Tris-HCl pH 8.0 and 200 mM NaCl and centrifuged at 17 500 rpm for 1 h at 4°C. The supernatant was incubated with Talon Affinity resin (Clonetech). To release the his-tagged complex from the Talon resin, the sample was treated with 3C protease overnight at 4°C. The next day, ion exchange chromatography was performed with a HiTrap Q HP column (GE Healthcare) using a gradient of NaCl from 50 mM to 1 M NaCl to remove bound nucleic acids. The sample was then further purified by size exclusion chromatography in 10 mM Tris HCl pH 8.0, 200 mM NaCl and 0.5 mM TCEP on a 16/60 Superdex 200 gel filtration column (GE Healthcare). Escherichia coli TadA was produced and purified using the same protocol as for the ADAT complex.

Ramos-Morales E., Bayam E., Del-Pozo-Rodríguez J., Salinas-Giegé T., Marek M., Tilly P., Wolff P., Troesch E., Ennifar E., Drouard L., Godin J.D, & Romier C. (2021). The structure of the mouse ADAT2/ADAT3 complex reveals the molecular basis for mammalian tRNA wobble adenosine-to-inosine deamination. Nucleic Acids Research, 49(11), 6529-6548.

+ Open protocol

+ Expand

Purification of Lectin Binding Proteins

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

Ultrapure guanidinium chloride (GdnCl) and ammonium sulfate were purchased from Calbiochem. Concanavalin A sepharose, HiTrap Q HP column, Superdex 75 pg column and gel filtration molecular mass standards were purchased from GE Healthcare Biosciences Ltd. Biogel P-100 was purchased from BioRad. A protein molecular mass marker set was purchased from Fermentas.

Biswas H, & Chattopadhyaya R. (2014). Thermal, Chemical and pH Induced Unfolding of Turmeric Root Lectin: Modes of Denaturation. PLoS ONE, 9(8), e103579.

+ Open protocol

+ Expand

Production and Purification of Recombinant G53A TTR

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

The recombinant human G53A TTR was produced using BL21(DE3) expression system using the pMMHa plasmid as described previously.^{20 (link)} Briefly, an isolated colony from the LB agar plate was inoculated into five milliliters of LB media and the small culture was added to the larger culture (0.5 L) after 4 hours of incubation. Protein expression was induced at an OD₆₀₀ of 0.6-0.7 with 1 mM IPTG and the bacteria cells were grown overnight (12-14 hrs) at 25 °C. The cells were harvested by centrifugation at 8000 rpm at 4 °C and the bacterial pellet was stored at −80 °C for at least 1 hr. The pellet was then resuspended in lysis buffer (10 mM Tris buffer, 150 mM NaCl, pH 8) and sonicated to disrupt the cells. After the sonication, supernatant was collected by centrifugation and ammonium sulphate was added (up to 50 %) to precipitate out other cellular proteins. The precipitates were removed by centrifugation, and the supernatant was dialyzed against 20 mM Tris buffer (1 mM EDTA and 1mM PMSF, pH 8) overnight at 4 °C. The G53A TTR was further purified with HiTrap Q HP column (GE Healthcare) followed by size exclusion gel chromatography using HiLoad 16/60 Superdex 200 column (GE Healthcare). The protein concentration was calculated using an extinction coefficient of 7.76 × 10^{4 (link)} M⁻¹ cm⁻¹ at 280 nm.

Dasari A.K., Arreola J., Michael B., Griffin R.G., Kelly J.W, & Lim K.H. (2020). Disruption of the CD loop by Enzymatic Cleavage Promotes the Formation of Toxic Transthyretin Oligomers through a Common Transthyretin Misfolding Pathway. Biochemistry, 59(25), 2319-2327.

+ Open protocol

+ Expand

Purification and Structural Analysis of HsCPAP Complexes

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

The rest of the protein samples eluted from IMAC were pooled and dialyzed for 3 h at 4°C against buffer H-150 (20 mM Hepes pH 7.4, 150 mM NaCl, 0.25 mM TCEP and 10% Glycerol). During the dialysis process, HsCPAP^897−1338 formed thin fiber-like structures that could be detected at plain sight. NS-EM visualization of the sample before and after dialysis showed a heterogeneous set of particles in both cases; interestingly, only in the last condition, modular rope-like structures of variable length could be seen. In an attempt to obtain fractions enriched with the elongated supramolecular complexes, the dialyzed sample was then applied on an anion exchange HiTrap Q HP column (GE Healthcare Life Sciences) equilibrated in the same buffer. The collected fractions were analyzed by NS-EM.

Alvarez-Cabrera A.L., Delgado S., Gil-Carton D., Mortuza G.B., Montoya G., Sorzano C.O., Tang T.K, & Carazo J.M. (2017). Electron Microscopy Structural Insights into CPAP Oligomeric Behavior: A Plausible Assembly Process of a Supramolecular Scaffold of the Centrosome. Frontiers in Molecular Biosciences, 4, 17.

+ Open protocol

+ Expand

Purification of GINS Complex

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

Sld5, Psf1, Psf3, and Psf2-3C-6xHis-Flag were overexpressed from ySK136. Cells were resuspended in buffer H, 1 M sorbitol, 0.02% NP-40, 2 mM ATP, and 0.5 M KCl. After lysis, the cleared lysate was diluted to 0.3 M KCl with buffer H. The lysate was then incubated with 1.5 mL of anti-Flag M2 affinity gel equilibrated with buffer M. The resin was washed with 20 CV of buffer M followed by 10 CV of buffer H, 0.02% NP-40, and 0.1 M KCl. GINS was eluted in the previous buffer + 3xFlag peptide. The Flag tag on Psf2 was removed with an overnight incubation (16 h) with HRV 3C protease. GINS was flowed over Complete His tag resin to remove uncut GINS and HRV 3C protease before applying the flow-through to a 1-mL HiTrap Q HP column (GE healthcare). GINS was eluted with a 20 CV gradient of 0.1–1 M KCl in buffer H and 0.02% NP-40. The peak fractions were dialyzed against buffer D.

Lõoke M., Maloney M.F, & Bell S.P. (2017). Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase. Genes & Development, 31(3), 291-305.

+ Open protocol

+ Expand

Purification of Histidine-Tagged Protein

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

The cell pellets were re-suspended in 20 mM Tris-HCl buffer (pH 7.0) with 500 mM NaCl (pH8.0) followed by sonication. The cell lysate was centrifuged at 10,000 × g at 4 °C for 20 min and the supernatant was applied to a nickel-charged resin column (Takara, Shiga, Japan). The column was washed with gradual washing containing 20–50–200–500 mM imidazole. The fraction eluted with 200 mM imidazole was collected and dialyzed against 20 mM Tris-HCl buffer (pH 7.4). After centrifugation the enzyme solution was loaded onto a pre-equilibrated HiTrap Q HP column (GE Healthcare) and the elution was performed with a linear gradient of NaCl from 100 to 500 mM. Under these conditions, most target protein was in the fractions of 200 mM NaCl eluate. The purity of each fraction was assessed by SDS-PAGE^{39 (link)}. Only those fractions showing a single band were pooled and the concentration of the resulting purified sample was determined to be 1.40 mg/ml using the method of Bradford with bovine serum albumin as a standard^{40 (link)}.

Wu Y., Mao G., Fan H., Song A., Zhang Y.H, & Chen H. (2017). Biochemical properties of GH94 cellodextrin phosphorylase THA_1941 from a thermophilic eubacterium Thermosipho africanus TCF52B with cellobiose phosphorylase activity. Scientific Reports, 7, 4849.

+ Open protocol

+ Expand

Purification of Cre and Cas9 Fusion Proteins

Cited 1 time

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

IL-31_K138ASNAP, NGF_R121WSNAP and the SNAP-tag were produced as previously described^{14 (link),15 (link),41 (link)}. cDNAs for CRE recombinase or Cas9 endonuclease together with a CLIP tag were cloned into a pETM-11 vector after a SenP2 cleavage site and expressed in E. Coli as fusion protein, carrying a His6-tag-SUMO at the N-terminus. His6-SUMO-CLIP-Cre and His6-SUMO-CLIP-Cas9 constructs were recovered from cell lysate with Ni-NTA resin (Qiagen, #30210) and eluted with 250 mM imidazole. An overnight (O/N) digestion with His-tagged SenP2 protease was performed at 4 °C in order to remove the SUMO tag-protein. Excess of imidazole was removed from solution through dialysis. Digested products were incubated again with Ni-NTA resin at room temperature (RT) for 1 hr. The flow-through containing only the protein of interest was collected. Further purification was performed with Ion Exchange chromatography in a HiTrapQ HP column (GE Healthcare) and finally with size exclusion chromatography in a Superdex 75 26/60 column (GE Healthcare). The protein-containing fractions were pooled and dialysed O/N against Phosphate-buffered saline (PBS), 1 mM DTT, 5% glycerol. Dialysed proteins were then concentrated (Amicon Ultra 10 kDa, Merck-Millipore), aliquoted, snap-frozen and stored at −80 °C.

Maffei M., Morelli C., Graham E., Patriarca S., Donzelli L., Doleschall B., de Castro Reis F., Nocchi L., Chadick C.H., Reymond L., Corrêa IR J.r., Johnsson K., Hackett J.A, & Heppenstall P.A. (2019). A ligand-based system for receptor-specific delivery of proteins. Scientific Reports, 9, 19214.

+ 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!