Neutrophils play a crucial role in autoimmune diseases beyond their well-characterized effector functions like phagocytosis and NETosis. Comprehensive assessment of neutrophil function is essential for understanding their diverse contributions to autoimmunity, including their roles in cytokine production and reactive oxygen species (ROS) generation. This chapter provides detailed methodologies for evaluating neutrophil immunomodulatory functions relevant to autoimmune pathology. We describe assays for quantifying cytokine and chemokine profiling by multiplex approaches and ROS production using flow cytometry. These methods enable researchers to dissect the nuanced contributions of neutrophils to autoimmune diseases, facilitating the identification of novel therapeutic targets.