AB-stained mucin bands were excised and subjected to reductive β-elimination, as described previously [12 (link), 13 (link)]. Briefly, the excised bands were incubated in 40 μL of 500 mM NaBH4 (191–11452; Fujifilm Wako Pure Chemical Corporation) in 50 mM NaOH (197–14891; Fujifilm Wako Pure Chemical Corporation) at 45°C for 16 h and the reaction was subsequently quenched by the addition of 8 μL of glacial acetic acid (017–00256; Fujifilm Wako Pure Chemical Corporation). The solutions were desalinated using a cation-exchange solid phase extraction cartridge (Oasis MCX, 60 mg; Waters Corporation, Milford, MA, USA) and concentrated and co-evaporated using 1% acetic acid in methanol to remove boric acid. The resulting residues were permethylated using iodomethane (137–02663; Fujifilm Wako Pure Chemical Corporation) and NaOH, desalinated, and dried as previously reported [12 (link), 13 (link)].
Acetic acid
Manufactured by Waters Corporation
Sourced in United States
Acetic acid is a colorless, transparent, and volatile liquid used as a chemical reagent in various laboratory applications. It has a distinctive sharp and pungent odor. Acetic acid is commonly used as a solvent, a pH adjustment agent, and a reagent in analytical procedures.
Automatically generated - may contain errors
Lab products found in correlation
Sodium hydroxide (naoh), by Fujifilm (1 mentions)
Iodomethane, by Fujifilm (1 mentions)
Glacial acetic acid, by Fujifilm (1 mentions)
Nabh4, by Fujifilm (1 mentions)
616 pump, by Waters Corporation (1 mentions)
Agilent hi plex h, by Agilent Technologies (1 mentions)
Af degasser, by Waters Corporation (1 mentions)
0.003 m h 2 so 4, by Merck Group (1 mentions)
Nanosep3k omega, by Pall Corporation (1 mentions)
Waters hplc system, by Waters Corporation (1 mentions)
717 autosampler, by Waters Corporation (1 mentions)
600 controller, by Waters Corporation (1 mentions)
Acetic acid, by Merck Group (1 mentions)
2 protocols using acetic acid
1
Reductive β-Elimination and Permethylation of Mucin
2
HPLC-RI Analysis of Acetic Acid
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Acetic acid concentrations of the autoclaved medium were determined using HPLC-RI.
Determination was performed using HPLC Waters system (Waters, Milford, USA) consisting of a Waters 600 controller, a Waters 2414 refractive index detector (HPLC-RI), a Waters 616 pump, a Waters 717 autosampler and a Waters 600 column oven.
Solvent was degassed in line using a Waters AF degasser. Separations were carried out on an Agilent Hi-plex H (7.7 × 300 mm Ø, particle size 8 μm) column (Agilent Technologies, USA) at a temperature of 65°C and a flow rate of 0.6 mL/min. The detector was set at 50°C. The mobile phase was 100% 0.003 M H2SO4 (Sigma-Aldrich).
Calibration has been performed using Acetic acid (>99.5%, J.T.Baker) at concentration levels of 50, 100, 200, 500, 1 000, 2 000 and 5 000 ppm. Data reprocessing was done using Empower 2 software. Prior to HPLC analysis, proteins were removed using Nanosep 3K Omega centrifugal devices (Pall Life Sciences, USA). After centrifugation at 11 000 × g for 5 min, the samples were ready for injection into the HPLC system.
Determination was performed using HPLC Waters system (Waters, Milford, USA) consisting of a Waters 600 controller, a Waters 2414 refractive index detector (HPLC-RI), a Waters 616 pump, a Waters 717 autosampler and a Waters 600 column oven.
Solvent was degassed in line using a Waters AF degasser. Separations were carried out on an Agilent Hi-plex H (7.7 × 300 mm Ø, particle size 8 μm) column (Agilent Technologies, USA) at a temperature of 65°C and a flow rate of 0.6 mL/min. The detector was set at 50°C. The mobile phase was 100% 0.003 M H2SO4 (Sigma-Aldrich).
Calibration has been performed using Acetic acid (>99.5%, J.T.Baker) at concentration levels of 50, 100, 200, 500, 1 000, 2 000 and 5 000 ppm. Data reprocessing was done using Empower 2 software. Prior to HPLC analysis, proteins were removed using Nanosep 3K Omega centrifugal devices (Pall Life Sciences, USA). After centrifugation at 11 000 × g for 5 min, the samples were ready for injection into the HPLC system.
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