Milk fat globule membrane (MFGM) material isolated from reconstituted buttermilk by microfiltration (i.e., whole MFGM) contains two major fractions, namely proteins (consisting mainly of MFGM-specific proteins and serum proteins) and lipids (original membrane polar lipids and contaminating triglycerides from the globule core). In this study, MFGM proteins and polar lipid (PL) concentrate were separated from whole MFGM material using solvent extraction. The particle size distribution, stability, surface protein and polar lipid load of oil-in-water emulsions prepared with protein or PL concentrate, individually or in combination, at various concentrations were examined. At low emulsifier concentrations (<2.3%), there was an interacting effect between proteins and PLs on the droplet size. The phase separation of emulsions prepared with a combination of 0.3% proteins and 0.3% PLs was similar to that of emulsions containing 0.3% proteins. The proteins were more preferentially adsorbed at the emulsion droplet surface compared with PLs.