Serum neutrophil gelatinase-associated lipocalin (sNGAL) is a marker of renal injury, and its concentrations are affected by inflammation. Therefore, it could serve as a useful biomarker of disease or fitness in high-level competition. However, it has not yet been determined if sNGAL concentrations are affected by exercise. The aim of this study was to determine whether concentrations of equine sNGAL were affected by 1000 m galloping as the form of exercise used in the study. Pre- and post-gallop sNGAL, serum amyloid A, and creatinine concentrations were evaluated in 14 healthy Thoroughbred racehorses. The results showed that short, high-intensity exercise did not significantly affect sNGAL concentrations in healthy horses (P =.42), and no significant difference was found in either creatinine or serum amyloid A before and after galloping (P >.05). Therefore, it was determined that sNGAL was not influenced by the type of exercise used in the study and could have the potential to be used as a routine laboratory screening tool in horses even after strenuous exercise. Future research should clarify its use in a larger population and a broader range of equine sport disciplines, including endurance-related exercise.