Peg 2000

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

PEG-2000 is a water-soluble polyethylene glycol polymer with an average molecular weight of 2,000 Daltons. It is commonly used as a reagent in various laboratory applications.

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4 protocols using peg 2000

Protein Purification Buffer Formulation

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Sarcosine, PEG400, PEG2000 (Alfa Aesar), Ficoll (GE Healthcare), guanidine hydrochloride (Thermo Fisher Scientific), ethylene glycol, glycine, potassium chloride, sodium chloride, sucrose and urea (Fisher Scientific) were used without further purification. Stock solutions were made by mixing the solute with 20 mM sodium phosphate buffer, pH 7.4, with the addition of 100 mM NaCl except for experiments where the concentration of NaCl or KCl was varied, which began free of additional salt. The same buffer was used for all dilutions.

Moses D., Guadalupe K., Yu F., Flores E., Perez A.R., McAnelly R., Shamoon N.M., Kaur G., Cuevas-Zepeda E., Merg A.D., Martin E.W., Holehouse A.S, & Sukenik S. (2024). Structural biases in disordered proteins are prevalent in the cell. Nature Structural & Molecular Biology, 31(2), 283-292.

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Synthesis of Ammonium-Based Ionic Liquids

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The PILs used in this work for the formation of ABS were synthetized in our laboratory according to the procedure previously described by McCrary et al.29 For the synthesis of PILs with amine-based cations with different carboxylate anions the following compounds were used: N-propylamine (98%); N-butylamine (99.5%); N-hexylamine (99.5%); N-octylamine (99%); butyric acid (≥ 99%); propanoic acid (≥ 99%); and hexanoic acid (≥ 99%). For the synthesis of [NH₄][But], ammonia (25% in water) from ChemLab and butyric acid (> 99%) were used. All compounds were purchased from Sigma-Aldrich (St. Louis, MO, USA) as colorless liquids and used as received. Triethylamine (99%) and dipropylamine (99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA), and were further distilled under vacuum prior to use to obtain colorless liquids. Glacial acetic acid (99.7%) was purchased from Macron (Center Valley, PA, USA) as a clear liquid and used as received. For comparison purposes [NH₄][OAc] (98%) was also used, which was supplied by Sigma Aldrich (St. Louis, MO, USA).
PEGs with average molecular weights of 600, 1000 and 3400 g mol⁻¹ (abbreviated as PEG-600, PEG-1000 and PEG-3400) were acquired from Sigma-Aldrich (St. Louis, MO, USA), while the one with an average molecular weight of 2000 g mol⁻¹ (PEG-2000) was purchased from Alfa Aesar. Double distilled water was used in all experiments.

Cláudio A.F., Pereira J.F., McCrary P.D., Freire M.G., Coutinho J.A, & Rogers R.D. (2016). A critical assessment of the mechanisms governing the formation of aqueous biphasic systems composed of protic ionic liquids and polyethylene glycol. Physical chemistry chemical physics : PCCP, 18(43), 30009-30019.

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Synthesis of MoO3 Nanoparticles

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MoO₃ nanoparticles were synthesized as follows: 2 g of ammonium heptamolybdate tetrahydrate (≥99.0%, Sigma-Aldrich, Saint Louis, MO, USA) was dissolved in a 50 mL mixture of H₂O and ethanol. An amount of 0.5 g of polyethylene glycol (PEG-2000, Alfa Aeser, Royston, UK) was then added to the mixture and stirred for 2 h. After the pH value was adjusted to 4.5 with an HNO₃ (65.0~68.0%, analytic reagent, Damao Chemical Reagent Factory, Tianjin, China) solution, the mixture was transferred to a Teflon-lined stainless autoclave and heated at 110 °C for 24 h. The sediment was filtrated, washed with H₂O, and dried overnight in a vacuum oven at 80 °C to obtain nano-MoO₃, which was designated as MoO₃(N).

Hu J., Liu J., Liu J., Li Y., Li P., Wang Y., Guan J, & Kan Q. (2020). Enhancing Methane Aromatization Performance by Reducing the Particle Size of Molybdenum Oxide. Nanomaterials, 10(10), 1991.

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NMR Analysis of NS4A Peptide

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The high-performance liquid chromatography-grade peptide with the sequence of KKGG STWVLVGGVLAALAAYCLTTGS GGKK (N → C), mimicking the N-terminal tail of NS4A, was purchased from Lipopharm.pl (Gdańsk, Poland). The flanking KKGG were added as solubility tags. PEG 600 and PEG 2000 were purchased from Alfa Aesar (Haverhill, MA), and Ficoll 400 from Sigma Aldrich (St. Louis, MO). CD spectra were recorded in water, in 10 mM phosphate buffer (pH 7.0), and in the presence of PEG 600, PEG 2000, or Ficoll 400 suspended in the phosphate buffer. In all CD experiments, the peptide concentration was 50 μM. The concentration of PEG crowders was 50 mg/mL and of Ficoll was 25 mg/mL because of problems with dissolving the peptide in Ficoll crowders. The CD spectra were collected using the Biokine MOS-450/AF-CD spectrometer with the Xe lamp. The acquisition time was 2 s with a resolution of 1 nm. Measurements were performed using a 0.1 cm cell, in the wavelength range 190–260 nm and at room temperature, with the high-tension values below 600 V. The presented CD spectra are the averages of three scans. Each biological experiment was conducted twice. The Savitzky-Golay (50 ) method was used to smooth the graphs.

Ostrowska N., Feig M, & Trylska J. (2021). Crowding affects structural dynamics and contributes to membrane association of the NS3/4A complex. Biophysical Journal, 120(17), 3795-3806.

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