Mass spectrometry analysis was carried out for the soluble fraction from all 14 donors and for the mucus fraction from a subset of six donors. Samples were first resuspended with 10 μL of 3% Acetonitrile + 0.1% formic acid and the total sample was analyzed using a nanoAdvanced UPLC (Bruker) with a 15 cm × 100 μm ProntoSil C18AQ column and 2 cm trap column (nanoLCMS Solutions). Mobile phase was H2O + 0.1% FA (A) and acetonitrile + 0.1% FA (B), peptides were separated using a gradient of 2%–40% B over 30 minutes 2 at a flow rate of 800 nL/minute and with the column temperature kept constant at 40°C. The column was connected through a Captive Spray nano source to an Impact Q-tof (Bruker). Data were collected over a mass range of 150–2200 m/z using a 1Hz MS scan and a total duty cycle time of 2 s, a setting that allows the Impact to adjust the acquisition time for each fragment scan based on precursor intensity while still collecting data on as many precursors as possible without exceeding the 2 s duty cycle. Data were processed using DataAnalysis 4.2 (Bruker), compounds were searched against the SwissProt database using Mascot 2.4 (Matrix Science) with the percolator algorithm, and protein and peptide results were assessed and filtered with ProteinScape 3.0 (Bruker).
Nanoadvanced uplc
Manufactured by Bruker
The NanoAdvanced UPLC is a liquid chromatography system designed for high-performance, high-resolution separation of complex samples. It features advanced fluidics and control systems to provide precise and reproducible results.
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Lab products found in correlation
2 protocols using nanoadvanced uplc
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Proteomic Profiling of Biological Fluids
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Proteomic Profiling of Biological Fluids
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Mass spectrometry analysis was carried out for the soluble fraction from all 14 donors and for the mucus fraction from a subset of six donors. Samples were first resuspended with 10 μL of 3% Acetonitrile + 0.1% formic acid and the total sample was analyzed using a nanoAdvanced UPLC (Bruker) with a 15 cm × 100 μm ProntoSil C18AQ column and 2 cm trap column (nanoLCMS Solutions). Mobile phase was H2O + 0.1% FA (A) and acetonitrile + 0.1% FA (B), peptides were separated using a gradient of 2%–40% B over 30 minutes 2 at a flow rate of 800 nL/minute and with the column temperature kept constant at 40°C. The column was connected through a Captive Spray nano source to an Impact Q-tof (Bruker). Data were collected over a mass range of 150–2200 m/z using a 1Hz MS scan and a total duty cycle time of 2 s, a setting that allows the Impact to adjust the acquisition time for each fragment scan based on precursor intensity while still collecting data on as many precursors as possible without exceeding the 2 s duty cycle. Data were processed using DataAnalysis 4.2 (Bruker), compounds were searched against the SwissProt database using Mascot 2.4 (Matrix Science) with the percolator algorithm, and protein and peptide results were assessed and filtered with ProteinScape 3.0 (Bruker).
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