Abstract: Background and Objective: Aloe dhufarensis is a local endemic medicinal herb used for centuries to treat diabetes, headache, fever, skin disorders and wound healing by the local people of Oman. The present research was initiated to substantiate the ethno-medicinal uses of this plant by assessing the phytochemicals and evaluating the antioxidant, anti-inflammatory and anti-microbial activityof Aloe dhufarensis. Materials and Methods: Dried, finely grinded leaves of A. dhufarensis were extracted with methanol for phytochemical analysis using standard protocols and several phytochemical such as saponins, alkaloids, terpenoids, flavonoids, steroids, tannins, glycosides, anthraquinones, phytosterol, coumarins and carbohydrates were detected in the extract. Results: Extracts displayed significant antioxidant property, exhibiting a potent DPPH scavenging activity with IC50 value of 83.46 μg mL–1 and a strong hydrogen peroxide scavenging activity with an IC50 value of 289.786 μg mL–1. Substantial inhibition of protein denaturation and anti-proteinase activity were recorded. Considerable antibacterial activity was observed against all 4 studied strains with the highest antibacterial activity recorded against the E. coli. Conclusion: The results reveal that A. dhufarensis possess a wide spectrum of phytochemicals that have significant pharmacological properties and substantiate the support to the scientific basis for the traditional use of this ethnomedicinal herb.