Prepacked columns

Manufactured by GE Healthcare

Prepacked columns are a type of laboratory equipment used for various chromatographic separation techniques. They contain a stationary phase material packed within a column, which allows for the separation and purification of complex mixtures. These columns can be used in a range of applications, such as ion exchange, size exclusion, and affinity chromatography, depending on the specific stationary phase and column characteristics.

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Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Surebeads protein a magnetic beads, by Bio-Rad (1 mentions) Akta chromatography system, by GE Healthcare (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions)

4 protocols using prepacked columns

Recombinant hFasLECD Protein Preparation

Cited 1 time

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hFasLECD containing single deletion mutation from 103 to 138 and double substitution mutations (N184Q and N250Q) [hFasLECD (139-281, N184Q, N250Q)] with an N-terminal FLAG-(GlyCysGlyGlyGlyGly) tag sequence (NFG1CG4-hFasLECD) and that with an N-terminal FLAG-(Gly)₅ tag sequence (NFG5-hFasLECD) were prepared as described (Muraki 2008 (link), 2014b (link)). FL-5Mal was obtained from Tokyo Chemical Ind. Dimethyl Sulfoxide, Super Dehydrated (Dry DMSO), l-Cysteine hydrochloride monohydrate and pre-cast gels for SDS-PAGE analysis (Supersep Ace, 10–20 % gradient gels) were from Wako Pure Chemicals Ind. BCA protein assay kit and Tris-(2-carboxyethyl)phosphine (TCEP) neutral pH solution were purchased from Thermo Fisher Scientific. Ion-exchange chromatography and size-exclusion chromatography were performed using prepacked columns from GE healthcare. Ultracentrifugation devices for sample concentration [Amicon Ultra 4 and 15, molecular-weight cut off (MWCO): 10 kDa] were supplied from Merck Millipore. Phosphate, acetate and Tris-hydrochloride buffers were the products of Nakarai Tesque (the former two) and Nippon Gene, respectively. SureBeads Protein A Magnetic Beads and washing buffer reagents used in immunoprecipitation experiments were from Bio-Rad Laboratories and Roche Diagnostics. Chemical structure of FL-5Mal was drawn using ChemBioDraw Ultra, ver. 14.

, & Muraki M. (2016). Preparation of a functional fluorescent human Fas ligand extracellular domain derivative using a three-dimensional structure guided site-specific fluorochrome conjugation. SpringerPlus, 5(1), 997.

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Methanosarcina acetivorans Protein Purification

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Acetate-grown Methanosarcina acetivorans was mass cultured and harvested as previously described (53 (link)). E. coli strain BL21(DE3) ΔiscR was a gift from J. Golbeck. HSCoB and CoMS-SCoB was a gift from T. Wood. All chromatography columns, resins, and prepacked columns were purchased from GE Healthcare. Purification of F₄₂₀ from methanol-grown M. acetivorans cells and preparation of F₄₂₀H₂ was performed as described elsewhere (54 (link)). All other chemicals were purchased from Sigma-Aldrich or VWR International.

Yan Z., Wang M, & Ferry J.G. (2017). A Ferredoxin- and F420H2-Dependent, Electron-Bifurcating, Heterodisulfide Reductase with Homologs in the Domains Bacteria and Archaea. mBio, 8(1), e02285-16.

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Albumin Depletion and Serum Fractionation

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All chromatography steps were carried out on an Äktapurifier Workstation from GE with prepacked columns from GE Healthcare. For removal of albumin serum from severely injured patients was passed manually over a Cibacron Blue affinity column (5 ml HiTrap Blue). Albumin and the bound nonalbumin proteins were eluted with 2 M NaCl in 20 mM sodium phosphate (pH 7.2). The albumin-depleted serum was collected and separately stored at -80°C until further processing.
Analytical size exclusion chromatography was achieved by fractionating albumin-depleted serum (150 μg/100 μl) on a Superose 6 10/300 GL gel filtration column. After chromatographic processing the serum fractions were stored at -80°C until further processing.

Sarabhai T., Peter C., Bär A.K., Windolf J., Relja B., Wesselborg S., Wahlers T, & Paunel-Görgülü A. (2017). Serum α-1 Antitrypsin (AAT) antagonizes intrinsic apoptosis induction in neutrophils from patients with systemic inflammatory response syndrome. PLoS ONE, 12(5), e0177450.

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Multiprotein Complex Purification Protocol

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The purification steps for both POLA1–POLA2–PRIM1–PRIM2 and p50–Ctc1–Stn1–Ten1 were performed at 4°C using an AKTA chromatography system with prepacked columns (GE Healthcare), following the same protocol. Cells were suspended in buffer A [30 mM Tris-HCl (pH 7.5), 200 mM NaCl, 10% (v/v) glycerol, 1mM dithiothreitol (DTT), and 25 mM imidazole] supplemented with protease inhibitor cocktail (Sigma), lysed by sonication, and centrifuged at 34,000×g for one hour. The supernatant was applied onto a 5-ml HisTrap HP column pre-equilibrated in buffer A. The column was washed with buffer A and the complex was eluted with buffer B [30 mM Tris-HCl (pH 7.5), 1 M NaCl, 10% (v/v) glycerol, 1mM DTT, and 400 mM imidazole]. The protein complex was digested overnight with 0.2 mg/ml TEV protease and buffer-exchanged to buffer A. The digest was applied onto a 5-ml HisTrap HP column pre-equilibrated in buffer A. The target complex was isolated in the column flow-through, concentrated to 10 ml, and then applied onto a Superdex 200 gel filtration column pre-equilibrated in buffer C [25 mM Tris-HCl (pH 7.5), 150 mM NaCl, and 1mM DTT]. The complex was collected from peak fractions and analyzed by SDS polyacrylamide gel electrophoresis.

He Y., Song H., Chan H., Liu B., Wang Y., Susac L., Zhou Z.H, & Feigon J. (2022). Structure of Tetrahymena telomerase-bound CST with Polymerase α-Primase. Nature, 608(7924), 813-818.

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