Maximal exercise performance is a multifactorial process in which the cardiovascular component, the innervation of the musculature, and the contractile and metabolic properties of skeletal muscle all play key roles. Here, protocols are provided for assessment of maximal running capacity of mice on a treadmill, with a combination of short high‐intensity paradigms primarily intended to test for maximal power and cardiovascular function, and longer low‐intensity paradigms to assess endurance and oxidative metabolism in skeletal muscle. The coupling of treadmill running to indirect calorimetry, to correlate performance measurements to maximal oxygen consumption, is also described. Curr. Protoc. Mouse Biol. 1:141‐154. © 2011 by John Wiley & Sons, Inc.