Acute phase proteins (APP) are the foundation to the innate immune response and have been shown to be valuable biomarkers, as increases can occur with inflammation, infection, neoplasia, stress and trauma. In reptilian medicine, little is known about acute phase protein responses and if these protein levels can be used to distinguish health compromise within a wildlife population. The purpose of this study was to characterize the plasma concentrations of the acute phase proteins (haptoglobin and serum amyloid A [SAA]) and protein electrophoretic profiles in free ranging Eastern box turtles (Terrapene carolina carolina) and assess any possible correlations between varying age class (adult vs. juvenile), sex (male, female, or unknown), season (spring, summer, or fall), or geographical location. Blood samples were obtained from 324 Eastern box turtles (Terrapene carolina carolina) from 2010 to 2012 at three sites in Illinois and one site in Tennessee, USA. Reference ranges were created for each age class, sex, and location. Significant differences were noted with SAA (age class, sex, season, state), haptoglobin (age class, sex, state, IL location), TP (sex, season, state, IL location), albumin (age class, season, state, IL location), alpha-1 (sex, season, IL location), alpha-2 (sex, season, state, IL location), beta globulins (age class, sex, season, state, IL location), gamma globulins (sex, season, state, IL location). As a non-specific marker of inflammation, acute phase protein testing is a valuable tool for health assessment of wildlife populations and may prove useful as a method of health status surveillance in free-ranging reptiles.