The effect of different extraction procedures on functional properties of mucilage extracted from Plantago major seed were investigated using response surface methodology. Extraction at 75°C, using 1:60 water:seed ratio at pH 6.8 was the best condition for maximum yield (15.18%), emulsion stability (67.4%), foam stability (88.4%), solubility (97.36%) and water absorption capacity (39.74g/g) of mucilage. At this optimum point, PMSM had, on average 82.85% carbohydrate, 76.79 mgGAE/gdry total phenol content, 97.8mgg−1 total flavonoid content and 915.54μgml−1 antioxidant activity. The results indicated that PMSM had average molecular weight of 1.2×106Da. FTIR analysis demonstrated the presence of carboxyl, hydroxyl and methyl groups and glycoside bonds. The chain flexibility parameter, activation energy, ζ-potential and droplet size for PMSM were determined as 946.09, 0.78×107J/kgmol, 15.23mV (at neutral pH) and 448.56nm, respectively. Intrinsic viscosity for PMSM in deionized water was 14.24dlg−1.