Biochemical markers are commonly used for detection, diagnosis, and management of various diseases. Creatine kinase (CK) is an enzyme found in most tissues, and is the best known marker for the identification of acute myocardial infarction (AMI). We review the most common techniques used to quantify creatine kinase and its cardiac specific fraction, CK-MB. Electrophoresis, immunoinhibition, monoclonal immunoassay, and CK-MB isoform assays are considered for efficacy, sensitivity and specificity, and timeliness in the setting of acute non-traumatic chest pain. Limitations of the various techniques for identifying AMI are identified. Current recommendations for CK-MB panels, including combinations with other cardiac markers, are included.