Detection of recombinant and native irisin in different species with antibody B. A: Recombinant irisin (lanes 1–8: 4, 2, 1, 0.5, 0.25, 0.125, 0.062, and 0.031 ng) and native irisin in sera of human*, rat, and horse detected with antibody C. All sera were albumin- and IgG-depleted, deglycosylated and 80 μg protein were resolved per lane. *Human samples: HS 4b and HS 5b (12 w training; details in Supplementary Table S2). B: Recombinant irisin (lane 1); recombinant glycosylated irisin before (lane 2) and after (lane 3) deglycosylation. Bovine plasma was spiked with 50, 100, 200, and 400 ng/ml irisin, albumin- and IgG-depleted, deglycosylated and 120 μg protein were resolved (lanes 4–7) and detected with antibody B. C: Human*, rat, mouse (mou.), baboon (bab.), and bovine (bov.) plasma/serum samples were albumin- and IgG-depleted and deglycosylated and 120 μg total protein were resolved per lane. Native baboon plasma (2 μL with ∼100 μg protein) was applied to lane 9. Lane 1: 20 ng deglycosylated recombinant irisin. Detection with antibody B. *Human samples: HS 6a and HS 7a (12 w training + acute exercise; details in Supplementary Table S1). M: molecular weight marker. Regions of interest are marked by boxes.