Four years after EU laws made it illegal to discriminate against passengers on grounds of disability or "reduced mobility", the Commission published new guidelines to clarify rights.
The move has been deliberately timed ahead of the arrival in London for the Olympic Games of thousands of disabled Paralympians and spectators.
EU Transport Commissioner Siim Kallas said: "Dealing with disability in life is a tough enough challenge: things shouldn't get even tougher when you arrive at the airport."
The new guidelines cover travellers at all EU airports and the operations of EU carriers anywhere in the world. They also cover non-EU carriers within or leaving Europe.
They follow discussions with airlines and airport authorities, governments and consumer and disability groups, and will "provide a real added value to the increased travel activity by disabled persons and persons with reduced mobility expected for the Para-lympics", said a Commission statement.