The objective of the present investigation was to assess the antimicrobial potential of Delonix regia leaf extract fractions. The crude extracts (chloroform and methanolic) of the leaves of the plant were subjected to fractionation using n-hexane, ethylacetate and methanol. The fractions obtained (A-F) were tested for phytochemicals and the confirmation of the same were achieved by thin layer chromatographic analysis. The in vitro antimicrobial activity was done using cup and plate method using Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus niger. The maximum antibacterial activity was exhibited by fractions E and F at concentration of 400 µg while fractions A and D were significantly effective against the fungal strain. The findings of the study provide a scientific basis to further investigate the potential of the leaves of D. regia.