A radioimmunoassay, specific for the isoenzymes of creatine kinase containing the M subunit of the enzyme (MM and MB creatine kinase), was employed to determine total creatine kinase concentrations in serum and neuromuscular tissues independently of the state of activity of the enzyme. This technique provides a method for the detection of inactive enzyme, which could be produced by inhibitors of the enzyme or by mutations involving the active site of the enzyme. A series of experiments were carried out to compare the amount of creatine kinase in various samples as assessed by normal enzyme kinetic procedures and by radioimmunoassay. The two techniques yielded equivalent results in all situations tested. Samples included serum from normal subjects and subjects with genetic and acquired diseases of muscle and also extracts from skeletal and cardiac muscle. Small quantities of immunoreactive enzyme were found in nervous tissue and assessed in terms of the incidence of the M subunit.