Anti-Jo-1 antibody is a type of autoantibody directed against histidyl-tRNA synthetase, a protein found in the cell nucleus. It is part of a group known as extractable nuclear antigen (ENA) antibodies. Testing for anti-Jo-1 is primarily used in the diagnosis and classification of idiopathic inflammatory myopathies, particularly antisynthetase syndrome, which can involve muscle inflammation, interstitial lung disease, arthritis, Raynaud’s phenomenon, and mechanic’s hands. The presence of this antibody helps clinicians differentiate between types of autoimmune muscle diseases and guide management.