A canola protein isolate having a protein content of at least about 90 wt % (N×6.25), preferably at least about 100 wt %, and consisting predominantly of the 2S protein and substantially free from the 7S and 12S proteins is prepared. In one aspect, canola oil seed meal is extracted with aqueous protein solution at an elevated temperature to preferentially extract 2S protein from the meal to produce a canola protein solution containing predominantly 2S protein. The 2S canola protein is recovered as an isolate. In another aspect, the canola oil seed meal is extracted with aqueous saline solution to extract 2S, 7S and 12S proteins from the meal. The aqueous protein extract solution is heat treated at an elevated temperature to precipitate 7S and 12S proteins and leave a 2S protein solution from which the isolate may be recovered. In a further aspect, the aqueous protein solution is concentrated prior to the heat treatment.