Abstract: The present study has been under taken with an objective to determine the antibacterial, antioxidant and cytotoxic activity of the leaf extract Indigofera tinctoria. The selected medicinal plant was collected from near by region of Coimbatore. Antibacterial, activity was carried against gram positive and gram negative bacteria. Antioxidant property was determined both quantitatively and qualitatively. Determination of cytotoxic activity of leaf extract was carried out on lung cancer cell line. The compound present in the extract were identified by GC-MS analysis. The extract screened for photochemical analysis was found to contain bioactive compounds like flavonoids, saponins, tannins, steroidal terpens, phenols and anthroquinone. The leaf extract had shown the ability to inhibit the growth of gram positive bacteria namely Bacillus pumilus, Staphylococcus aureus and Streptococcus pyogenes and zone of inhibition of 16 and 17 mm, respectively but not shown growth of inhibition on gram negative bacteria Escherichia coli and Pseudomonas aeruginosa. Strong antioxidant activity was observed both qualitatively and quantitatively. The strong antioxidant was observed at 250 μg mL-1 with an IC50 value of 51.66 which is higher than that of standard ascorbic acid. The cytotoxic effect of leaf extract on lung cancer cell line NCI-H69 was studied. The percentage cell viability of cells was found to decrease at increasing concentration. GC-MS analysis of the leaf extract shows 6 compounds. This study suggests that ethanol extract of Indigofera tinctoria have profound antibacterial, antioxidant and cytotoxic effect.