The present study evaluates the physicochemical attributes, antibacterial efficacy, and antioxidant capacities of four distinct varieties of honey from the West Bank region of Palestine: Assal Barsem (Medicago sativa) AB, Assal Morar (Centaurea dumulosa Boiss) AM, Assal Horfesh (Silybum) AH, and Assal Sader (Ziziphus spina-christi) AS. The analysis encompassed parameters such as pH, electrical conductivity, Total Flavonoid Content (TFC), and Total Phenolic Content (TPC). Furthermore, the antioxidant potential was gauged through Total Antioxidant Capacity (TAC) determination and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. In addition, the antibacterial effectiveness of the honeys was measured against a spectrum of bacterial strains including Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Streptococcus pneumoniae, Klebsiella pneumoniae, Haemophilus influenzae, and Bacillus subtilis, utilizing minimum inhibitory concentration (MIC) and Minimum Bactericidal Concentrations (MBC). The outcomes of the physicochemical analysis adhered to the quality benchmarks outlined by the European Union Commission and the Codex Alimentarius Commission. The MIC and MBC values exhibited notable variance across the tested honey varieties, with MIC values ranging from 0.024% w/w to 1.56% w/w, and MBC values ranging from 0.048% w/w to 3.15% w/w. Particularly, AH demonstrated superior efficacy against all seven bacterial strains, with MIC values spanning from 0.1 to 0.6% w/w, and MBC values ranging from 0.3% w/w to 0.8% w/w. Staphylococcus aureus and Escherichia coli were notably susceptible to all honey samples. Collectively, our findings underscore the therapeutic potential of Palestinian honey varieties, highlighting their multifaceted health-promoting attributes. Further exploration is warranted to elucidate the mechanistic underpinnings of bioactive constituents and explore their potential applications in healthcare.