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"A lot of them are donating their stage tips, they're donating the money they made from private dances for the night".
Following digestion, peptides were desalted using C18STop And Go Extraction (STAGE) tips (made "in-house") [19].
The labeled peptides were then fractionated on C18-SCX-C18 C18-SCX-C18 C18-SCX-C18ep ammonium acetate elution gradient [21] and the dried peptide fractionSTAGEe resuspended in 1% tipslusingcetic acid, 3% acetonitrile, and 0.5% acetic acid.
Peptide digests were purified using the C18 flavor of STop And Go Extraction (STAGE) tips [46] and eluted peptides were dried and labelled by reductive dimethylation using formaldehyde isotopologues [22], [47].
Fractions eluted from the SCX column were desalted with C18 Empore (3 M, Minneapolis, MN, USA) stop and go extraction (STAGE) tips [54] and subsequently subjected to nano-flow RP-HPLC using the Ultimate LC system (Dionex) equipped with a nanoflow calibration cartridge at a flow rate of 250 nL/min.
The resulting peptides were desalted using Stage tips [ 60].
Samples were stored on STAGE tips at 4°C as needed.
The resulting peptide mixtures were extracted and desalted with stage tips (Thermo Scientific, USA).
Peptides were desalted with C18 stage tips (Thermo Scientific, West Palm Beach, FL) and dried under vacuum.
Enriched phosphopeptides were eluted and desalted by C18 columns (Stage tips).
The peptide samples were desalted using C18 stage tips [ 20] prior to nanoLC-MS/MS.
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