This study was conducted in an attempt to determine if the biomechanical and anatomical factors associated with iliotibial band (IT band) injury are quantitatively different between IT band injured and healthy runners. Positive correlation between the measurements and injury would provide information that could be used to develop better prevention and treatment programs for runners susceptible to this common injury. Forty runners, 20 injured and 20 healthy, participated in the study and were divided into 4 groups in order to examine the effects of leg length discrepancy. The 20 injured participants were randomly assigned into one of two groups of 10 (INJ-1 or INJ-2). INJ-1 was compared to 10 healthy control (CON) individuals. INJ-2 was compared to a group of 10 healthy runners (EXP) who trained for one week with a 1.27 cm felt heel pad in the shoe of their longer leg. Data were collected from all participants using a runner's questionnaire, 13 lower extremity anatomical measurements, 4 clinical lower extremity assessments, and 2D kinematics captured from the sagittal and frontal planes during treadmill running. Kinematic results indicated the INJ-1 group had a greater knee flexion angle than the CON group (p < 0.05). No other direct comparisons revealed statistically significant differences between groups, nor did a discriminant function analysis based on 9 anatomical measurements. Analysis of the running questionnaire responses did not indicate significant difference in training regimen between the injured or healthy participants. In summary, it was not possible to clearly identify differences in the biomechanical, anatomical, and training regimen factors most commonly thought to predispose individuals to iliotibial band injury. We believe that future research should focus on pelvic motion, pelvic muscle activity, leg strength, and/or flexibility variables.