Ninety-four blood samples were collected from 48 (29 males and 19 females) free-ranging Florida panthers (Felis concolor coryi) captured in southern Florida (USA) from 1983 to 1994 for routine hematological and serum biochemical analysis. Florida panthers in the northern portion of their range had significantly higher red blood cell (mean ± SD = 7.923 × 10⁶ ± 0.854 × 10⁶/μl), hemoglobin (12.53 ± 1.66 g/dl), and packed cell volume (36.97 ± 4.27%) values compared to those of panthers localized in more southern parts of Florida (7.148 × 10⁶ ± 1.045 × 10⁶/μl, 11.60 ± 1.62 g/dl, and 34.82 ± 5.99%, respectively). Adults had significantly higher mean serum total protein (7.50 ± 0.59 g/dl) and packed cell volume (36.90 ± 4.97%) values than juveniles (6.88 ± 0.49 g/dl and 34.54 ± 5.30%). However, mean serum albumin concentrations were significantly higher in juveniles (3.80 ± 0.26 g/dl) when compared to adult values (3.58 ± 0.26 g/dl). Mean serum calcium concentrations were significantly higher in juveniles (10.33 ± 0.39 mg/dl) than in adults (9.66 ± 0.45 mg/dl). Additionally, mean serum iron concentrations were significantly higher in those panthers of intergrade genetic stock compared to values in those of authentic genetic stock (105.6 ± 72.1 μg/dl versus 59.3 ± 19.7 μg/dl, respectively).