Barber had desperately tried to ready himself after suffering a left thigh injury on his final carry of the Cowboys' 33-31 loss to the New York Giants in Week 2.
Barber had limited participation in practice on Friday.
Defensive tackle Nick Hayden was back in the Carolina lineup after missing last week's loss to Atlanta with a bruised left big toe.
Owner Jerry Jones had described Barber's status as "doubtful" before the game.
Speaking to a group of reporters at a MNF event in Dallas, Jones said coaches would take a "conservative" approach to Barber playing because the Cowboys have two healthy backs in Felix Jones and Tashard Choice ready to play against the winless Panthers.
Information from ESPN.com NFC East blogger Matt Mosley and The Associated Press was used in this report.