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. 2020 Jan 31;34:124–135. doi: 10.1016/j.molmet.2020.01.016

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© 2020 The Author(s)

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Detection of irisin before and after deglycosylation. A: Deglycosylation of recombinant glycosylated irisin and human serum samples* with PNGase F or protein deglycosylation Mix II (NEB) and detection with antibody C. *Human samples: HS 2a, HS 3 (acute exercise; details in Supplementary Table S1). B: Detection of different amounts of recombinant glycosylated irisin (2, 1, 0.5, 0.25, 0.125, and 0.062 ng) with antibody B before (lanes 1–6, left panel) and after deglycosylation (lanes 1–6, right panel). Glycosylated recombinant irisin was added to albumin-depleted bovine plasma. Linearity of detection is shown in the right panel. Glycosylated irisin (left panel) could not be quantified (details in Supplementary Figure S2D). M: molecular weight marker. Regions of interest are marked by boxes.