Since the characterisation of Interleukin-6 as a myokine, the concept that skeletal muscle is a bona fide endocrine organ has gained considerable support. Here, we adopted the use of fully labelled ¹³C Lysine SILAC mice to screen skeletal muscle for myokine candidates. We pooled the gastrocnemius muscles from three, unlabelled C57/BL6 mice either exposed to 90 minute treadmill running or non-exercised controls and combined 1:1 with a spike-in SILAC gastrocnemius standard. After separation of 10 fractions by SDS-PAGE and trypsin/Lys-C enzymatic digestion, the resultant peptides were analysed by LC-MS/MS using an Orbitrap Elite. In all, we identified 2280 proteins with a 5% peptide false discovery rate. 593 of these proteins were quantified across exercise and control experiments, 337 of which contained at least two SILAC ratios. Enrichment of the data set for the gene ontology cellular component, extracellular region or space, distinguished a selection of candidate myokines such as Decorin, Annexin A2 and Mimecan that are currently undergoing validation.