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This kind of precipitation allows resuspension of the protein pellet in isoelectric focusing buffer contains 8 M urea, 2 M thiourea, 10 mM 1,4-dithiothreitol (DTT), 2 mM TCEP (Tris(2-carboxyethyl)-phosphine-hydrochloride), 1% (w/v) CHAPS, 1% (w/v) Triton X-100, 1% (w/v) amidosulfobetaine-14 (ASB), and 0.4% (v/v) ampholytes (pH 310)

In RA-FLS, an increase in glutamine transporters could support the role of glutamine as an alternate carbon source in the absence of glucose 45

electrochemiluminescence detection of each MSD SULFO-TAG was captured and quantified using SECTOR Imager instrumentation (MSD, Gaithersburg, MD, USA), according to manufacturers protocols as reported previously ( The raw data were analyzed using the Discovery Workbench 4.0 software (MSD) by fitting signal from the control calibration curves to a four-parameter logistic model and then back-fitting the electrochemiluminescence signal from each sample to calculate the unknown concentration

JCSM Rapid Communications (2020) 7