Steady-state fluorescence experiments were performed using an Agilent Cary Eclipse fluorescence spectrophotometer. Samples of various concentrations of PEG 20k and other variable-sized PEG molecules were prepared by dissolving the required quantity of PEG in water or water along with 100 mM NaCl and 30 mM phosphate buffer of pH 7.4 to mimic biological conditions. Solutions of 100 mg/mL PEG 20k in the presence of six different metal ions were prepared using 10 mM NaCl, MgCl2, CoCl2, FeCl3, CrCl3, and K2CrO4. Blank samples containing only water and the corresponding metal salt were prepared for each of the six ionic compounds. PEG 600, PEG 2k, and PEG 8k were purchased from Thermo Fisher Scientific and were used as received without further purification. PEG 20k was obtained from Thermo Fisher Scientific and MilliporeSigma. Fluorescence of PEG 20k was recorded before and after purification with diethyl ether. Samples were excited within a range of 210–300 nm, and the emission spectra were recorded from 300 to 400 nm with 5 mm spectral slits. All experiments were performed using a quartz cuvette with a 1 cm optical path length and were done in triplicate.
Peg 600
Manufactured by Thermo Fisher Scientific
Sourced in United States
PEG 600 is a type of polyethylene glycol (PEG) with an average molecular weight of 600 g/mol. PEG 600 is a colorless, odorless, and viscous liquid. It is commonly used as a solvent, plasticizer, and surfactant in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Cary eclipse, by Agilent Technologies (1 mentions)
Cell culture media, by Thermo Fisher Scientific (1 mentions)
Matrigel basement membrane matrix, by Corning (1 mentions)
Corn oil, by Fujifilm (1 mentions)
2 2 2 tribromoethanol, by Merck Group (1 mentions)
Polyethylene glycol peg 400, by Merck Group (1 mentions)
Cremophor el, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Solutol hs 15, by BASF (1 mentions)
Sodium heparin, by Leo pharma (1 mentions)
Peg 1000, by Thermo Fisher Scientific (1 mentions)
Fluorescein, by Merck Group (1 mentions)
Ammonium hydroxide nh4oh, by Thermo Fisher Scientific (1 mentions)
Acetic acid hoac, by Merck Group (1 mentions)
Peg200, by Thermo Fisher Scientific (1 mentions)
Gentamycin sulfate 600 iu mg, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by American Type Culture Collection (1 mentions)
9 protocols using peg 600
1
Fluorescence Characterization of PEG Compounds
2
Multiplex Assay for Cytokine Profiling
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The Flavin Adenine Nucleotide, Dicarboxylic Poly Ethylene Glycol (PEG)-600, cell culture media, fetal bovine serum (FBS), and penicillin-streptomycin were obtained from Thermo Fisher Scientific (Waltham, MA, USA). The 5-fluorouracil (5-FU) was purchased from Shanghai Xudong Haipu Pharmaceutical Co., Ltd. (Shanghai, People's Republic of China).
The Bio Legend LEGENDplexTM multiplex bead-based assay kit was from BioLegend (San Diego, CA, USA). The Matrigel® Basement Membrane Matrix was from Corning (Tewksbury, MA, USA). HEMAVET950FS animal blood analyzer special reagents were from Drew Scientific, Inc., USA. Isoflurane was obtained from RWD Life Science Co., Ltd. (Shenzhen, People's Republic of China).All other reagents were either obtained from Sigma-Aldrich (St. Louis, MO, USA) or noted otherwise.
The Bio Legend LEGENDplexTM multiplex bead-based assay kit was from BioLegend (San Diego, CA, USA). The Matrigel® Basement Membrane Matrix was from Corning (Tewksbury, MA, USA). HEMAVET950FS animal blood analyzer special reagents were from Drew Scientific, Inc., USA. Isoflurane was obtained from RWD Life Science Co., Ltd. (Shenzhen, People's Republic of China).All other reagents were either obtained from Sigma-Aldrich (St. Louis, MO, USA) or noted otherwise.
3
Preparation and Use of Avertin Solution
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C16 was prepared as described earlier [5 (link)]. A tribromoethanol injectable solution (Avertin) was prepared as described in Section E in S1 File . In-house prepared Milli-Q water was used. Cremophor EL (Sigma, Germany), corn oil (Wako, Japan), dimethyl sulfoxide (DMSO, Sigma-Aldrich, Germany) anhydrous N,N-dimethylacetamide (DMA, Wako, Japan), ethanol (Merck, Germany), Medigel Sucralose (2 oz cups, ClearH2O, USA), 2-methyl-2-butanol (amylene hydrate, Sigma-Aldrich, Germany) 1,2-propanediol (propylene glycol, PG) (Wako, Japan), polyethylene glycol (PEG) 400 (Sigma, U.S.A), PEG 600 (Alfa Aesar, Great Britain or Sigma, USA), sodium heparin (5000 units/mL injectable solution, LEO Pharma, Belgium) Solutol HS-15 (BASF, Germany) 2,2,2-tribromoethanol (Sigma-Aldrich, Germany) were used as obtained from the indicated suppliers.
4
Preparation of Organic Reagents and Solvents
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All applied organic reagents and solvents were of reagent grade purity and were used without further purification, unless stated otherwise. The 1-Methyl-1-phenylethyl hydroperoxide (cumyl hydroperoxide, CHP) was purified by the method described elsewhere [21 (link)].
The 1-Bromobutane (>98%) and cyclohexane were distilled prior to being used for synthesis. PEG (polyethylene glycol) 200, PEG 600, and PEG 1000 were purchased from Alfa Aesar GmbH & Co KG (Karlsruhe, Germany). PEG MM (polyethylene glycol monomethyl ether) 350, PEG MM 550, PEG MM 1100, PEG MM 2000, PEG DM polyethylene glycol dimethyl ether) 500, and PEG DM 1000 were purchased from Fluka Chemie GmbH (Steinheim, Germany).
The 1-Bromobutane (>98%) and cyclohexane were distilled prior to being used for synthesis. PEG (polyethylene glycol) 200, PEG 600, and PEG 1000 were purchased from Alfa Aesar GmbH & Co KG (Karlsruhe, Germany). PEG MM (polyethylene glycol monomethyl ether) 350, PEG MM 550, PEG MM 1100, PEG MM 2000, PEG DM polyethylene glycol dimethyl ether) 500, and PEG DM 1000 were purchased from Fluka Chemie GmbH (Steinheim, Germany).
5
Fluorescein-TEOS Silica Nanoparticles
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Tetraethyl orthosilicate (TEOS) and fluorescein were purchased from Sigma-Aldrich Chemical Co. (Milwaukee, WI, USA). Acetic acid (HOAc) was from EMD Millipore (Billerica, MA, USA), ammonium hydroxide (NH4OH) was from Fisher Chemical (Fair Lawn, NJ, USA), phosphate-buffered saline (PBS) was from ATCC (Manassas, VA, USA) and polyethylene glycol (PEG) with MW = 200 Da (PEG200), and MW = 600 Da (PEG600), respectively, were purchased from Alfa Aesar (Ward Hill, MA, USA). Gentamycin sulfate 600 IU/mg was from Alfa Aesar.
6
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.
7
Ionic Liquids as Pt Nanoparticle Substrates
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Three different ionic liquids, 1-ethyl-3-methylimidazolium
triflate (Emim Tf), 1-decyl-3-methylimidazolium triflate (Dmim Tf),
and 1-decyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide
(Dmim Tf2N), and the polymer polyethylene glycol 600 (PEG 600) were
used as liquid substrates in the present study. All ionic liquids
were more than 99% pure and purchased from Iolitec GmbH, and PEG 600
was purchased from Alfa Aesar and 95% pure. Ionic liquids representing
a range of alkyl chain lengths (Emim Tf and Dmim Tf) were chosen to
elucidate any Pt nanoparticle size dependence on the alkyl chain length.
Dmim Tf2N was chosen to compare two long-chained ionic liquids with
different anions.
triflate (Emim Tf), 1-decyl-3-methylimidazolium triflate (Dmim Tf),
and 1-decyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide
(Dmim Tf2N), and the polymer polyethylene glycol 600 (PEG 600) were
used as liquid substrates in the present study. All ionic liquids
were more than 99% pure and purchased from Iolitec GmbH, and PEG 600
was purchased from Alfa Aesar and 95% pure. Ionic liquids representing
a range of alkyl chain lengths (Emim Tf and Dmim Tf) were chosen to
elucidate any Pt nanoparticle size dependence on the alkyl chain length.
Dmim Tf2N was chosen to compare two long-chained ionic liquids with
different anions.
8
Cytokine Profiling: Optimizing Sample Preparation
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The 8-plex quality control mixture provided in the Bio-Plex Pro™ Human Cytokine 8-plex Assay (cat. M50000007A) has been diluted 1:1 in sterile PBS (Gibco, Thermo Fisher Scientific, Waltham, MA, USA). A volume of 60 µL has been taken for each of the following condition: UVC exposure for 10 and 60 mins as described in Section 2.2.2 or applied to Amicon® Ultra-100 centrifugal filter device (Cat UFC510096, Merck Millipore Ltd., Burlington, MA, USA) untreated or pre-treated with BSA (1 mg/mL) (Cat A4506, Sigma-Aldrich, Milan, Italy) or PEG600 (1 mg/mL) (Cat 81180 Fisher Scientific, Milan, Italy). After centrifugation at 3000 g for 30 min at 4 °C, the supernatant was collected from the sample reservoir with a pipette. After removal of any liquid left in the column, a volume of 60 µL has been applied to the filter for washing of the filter itself and then collected. All samples were run on the Bio-Plex200 System and on the 2100 Bioanalyzer as described in Section 2.5 and Section 2.6 , respectively. Three independent experiments have been performed.
9
DNA Folding Under Molecular Crowding
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Each CD sample, including controls,
was prepared with 5 μM of DNA in 1500 μL of Tris-KCl solution
(0.1 M KCl and 0.1 M Tris (pH 7.3)). If the complementary strand was
added, its concentration was also 5 μM. Three cosolutes, ethylene
glycol (EG), polyethylene glycol (PEG) 600, and PEG 8000 (Fisher)
were used to mimic naturally occurring biological crowders of different
sizes. Cosolutes were added at a w/v concentration of 10, 20, 30,
and 40%. Samples were annealed by heating to 95 °C followed by
gradual cooling to 25 °C over ∼3 h. When longer-term storage
was necessary, DNA solutions were stored in a specially dedicated
refrigerator at 8 °C. At this temperature, the DNA strands maintain
the same folded conformation as at 25 °C.
was prepared with 5 μM of DNA in 1500 μL of Tris-KCl solution
(0.1 M KCl and 0.1 M Tris (pH 7.3)). If the complementary strand was
added, its concentration was also 5 μM. Three cosolutes, ethylene
glycol (EG), polyethylene glycol (PEG) 600, and PEG 8000 (Fisher)
were used to mimic naturally occurring biological crowders of different
sizes. Cosolutes were added at a w/v concentration of 10, 20, 30,
and 40%. Samples were annealed by heating to 95 °C followed by
gradual cooling to 25 °C over ∼3 h. When longer-term storage
was necessary, DNA solutions were stored in a specially dedicated
refrigerator at 8 °C. At this temperature, the DNA strands maintain
the same folded conformation as at 25 °C.
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