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7 protocols using mesityl oxide

1

Polysaccharide-Based Biomaterial Synthesis

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PDDAC (20 wt% in water; M.W. 400,000–500,000), mesityl oxide (MO), sodium hydroxide, formic acid, tris(hydroxymethyl)aminomethane (Tris), d,l-Thi, d,l-Tri, d,l-Eth, d,l-Pro, d,l-Met, α-CD, β-CD, γ-CD, hydroxypropyl-α-CD (Hp-α-CD), Hp-β-CD, and Hp-γ-CD were obtained from Sigma–Aldrich Corporation (St. Louis, MO). The pH of formic acid was adjusted to 2.0–4.0 by adding 100 mM Tris. Water used in all experiments was doubly distilled and purified using a Milli-Q system (Millipore, Milford, MA).
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2

Synthesis and Characterization of PMAO

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Example 3

A 20 ml vial with a magnetic stir bar was charged with 3.5 g (48.6 mmol) TMAL (ex-AkzoNobel).

Mesityloxide (4-methylpent-3-en-2-one, ex-Sigma Aldrich; 3.30 g; 33.6 mmol; 0.69 eqv.) was then dosed over a period of 47 minutes. The reaction temperature was kept below 50° C. A clear and slightly viscous intermediate reaction mixture was obtained.

1 g of the prepared intermediate reaction mixture was charged to a 20 ml vial equipped with a magnetic stir bar. The vial was placed in a heating block and heated to 100° C. After about 8 minutes of heating at 100° C., a violent uncontrollable exothermic reaction occurred. All the liquid present in the reaction mixture was boiled off leaving a foamy substance in the vial.

In order to better control the exothermic reaction, a new 20 ml vial equipped with a magnetic stir bar was charged with 0.3 g of the prepared intermediate reaction mixture. The reaction mixture was diluted with 0.7 g TMAL and heated to 100° C. After 13 minutes, an exothermic, but controllable reaction occurred. Over a period of about 30 minutes, the remaining intermediate reaction mixture was dosed to the 20 ml vial. The reaction was exothermic and the reaction temperature was kept between 80 and 90° C. A clear, bright red, and slightly viscous liquid was obtained. The liquid solidified upon storage at −20° C.

The formation of PMAO was confirmed by 1H-NMR.

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3

Functionalized Carbon Nanotubes for Research

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Carbon nanotubes from NanoLab Inc (Waltham, MA) included precarboxylated carbon nanotubes (PD15L1–5-COOH) with outer diameter 15 ± 5 nm and length 1– 5 μm, and carbon nanotubes (PD15L5–20) with outer diameter 15 ± 5 nm and length 5– 20 μm. Precarboxylated carbon nanotubes (US4358) with outer diameter 10 – 20 nm and length 0.5– 2 μm were from US Research Nanomaterial, Inc (US-Nano, Houston, TX). Morpholinopropane-1-sulfonic acid (MOPS), methanol, sodium hydroxide and mesityl oxide were from Sigma-Aldrich Corp. (St. Louis, MO). MOPS (25 mM) was prepared in deionized water obtained from an Elga Purelab ultra water system (Lowell, MA), and the pH adjusted to 7 using sodium hydroxide. Acid treatment was accomplished with 95% sulfuric acid purchased from J.T. Baker (Center Valley, PA) and 69% Nitric Acid purchased from GFS Chemicals (Columbus, OH). The amidated peptide WRWWWW-NH2 was purchased from Bachem (Torrance, CA).
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4

ATV Characterization via Analytical Techniques

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The investigated sample of ATV was gifted from APL Research Centre Laboratories (a division of Aurobindo Pharma Ltd., Hyderabad.). Analytical reagent (AR grade) mesityl oxide, diacetone alcohol, o-xylene, methylene chloride, decane, ethanol, acetone, isopropyl alcohol (2-propanol), methyl tert-butyl ether, benzene, n-heptane, methyl isobutyl ketone (4-methyl-2-pentanone), toluene, and 1-methyl-2-pyrrolidinone were procured from Sigma-Aldrich (Steinheim, Germany). ACS grade formic acid was procured from Merck & HPLC water was procured from Merck, Mumbai, India.
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5

Synthesis and Analysis of Sulfur Compounds

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UPLC/MS grade water, methanol and formic acid (>99.98%) were purchased from Biosolve Chimie, Dieuze, France; deuterium (99.8% D), dithiotreitol (98%), (E)-2-hexenal (98%), 2-hexyn-1-ol (≥97%), Boc-Cys-OH (≥98.5%), (Boc-Cys-OH)2 (≥98%), ethanol-OD (≥99%), hydrochloric acid (37%), L-glutathione reduced (98%), Lindlar catalyst, manganese(IV) oxide (≥99%), mesityl oxide (≥90%), mesityl oxide d10 (>98% D), triethylamine (≥99%) were purchased from Sigma-Aldrich, St Quentin Fallavier, France, and H-Cys-Gly-OH (98%), H-Glu(Cys-OH)-OH (98%)from Bachem, Bubendorf, Suisse; triethylsilane (≥98%) was obtained from TCI Europe, Zwijndrecht, Belgium; while di-sodium hydrogen phosphate (≥99%), sodium carbonate (≥99%), sodium dihydrogen phosphate dihydrate (≥99%)and trifluoroacetic acid (≥99.9%) were purchased from Carl Roth, Karlsruhe, Germany.
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6

Optimized SDS-PAGE Analysis of Therapeutic Antibody

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Ultra-low gelling temperature agarose, sodium dodecyl sulfate (SDS), boric acid, Tris, EDTA·Na2, mesityl oxide, sodium hydroxide, hydrochloric acid, HPLC grade water, glacial acetic acid, glycerol and 2-mercaptoethanol were purchased from Sigma Aldrich (St. Louis, MO, USA). The 10 kDa size standard and the SDS-MW sample buffer were from Bio-Science Kft (Budapest, Hungary). The PNGase F enzyme was from the Bio-Nanosystems Laboratory, University of Pannonia (Hungary). The therapeutic monoclonal antibody drug (omalizumab, 10 mg/mL) was a kind gift of the Borsod Academic County Hospital (Miskolc, Hungary).
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7

Proteomics Sample Preparation Reagents

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Cytochrome c, myoglobin, ubiquitin, bovine serum albumin (BSA), hemoglobin, trypsin, mesityl oxide, diacetone alcohol, MS-grade trifluoroacetic acid (TFA), and formic acid were purchased from Sigma-Aldrich (Oakville, CA). Model peptides were purchased from GL Biochem Ltd. (Shanghai, China). HPLC-grade acetone, acetonitrile, and methanol were from Fisher Scientific (Ottawa, CA). Reagent-grade glacial acetic acid was from Caledon Laboratory Chemicals (Georgetown, CA), while reagent-grade ammonium hydroxide (28% solution) was from ACP Chemicals Inc. (Montreal, CA). Chemicals used for digestion, including urea, dithiothreitol (DTT), and iodoacetamide (IAA), were from Bio-Rad (Mississauga, CA). Throughout the study, Milli-Q purified water (18.2 MΩ cm) was used.
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