Mycobacterium habana, a cultivable nonpathogenic mycobacterium provides appreciable resistance in mouse against M. tuberculosis infection. This study is aimed at identification and characterization of protective antigens of M. habana. Protective potential of antigens of cell wall (CW), cell membrane (CM), cytosol (CS) and peripheral and integral compartments of the membrane fraction of M. habana was explored against experimental tuberculosis in mouse. Peripheral and integral membrane proteins were characterized by SDS-PAGE and differential staining with silver and periodic acid. Results reveal that protective antigens are distributed in both peripheral and integral membrane compartments though such effect is dominant in the former. Polysaccharide staining showed that LAM, LM and PIMs have a preference for the detergent phase. Peripheral and integral compartments constitute, respectively, 68 and 31% of the total membrane protein.