The National Football League requested the judge put her ruling to lift an owner-imposed labor lockout on hold.
U.S. District Court Judge Susan Nelson said Wednesday the NFL "has not met its burden for a stay pending appeal, expedited or otherwise."
On Monday, Nelson ruled for NFL players in an ongoing labor dispute between the athletes and team owners. Nelson granted an injunction to lift the lockout.
Players are seeking a new collective bargaining agreement with team owners. Some players filed an antitrust lawsuit against the owners.
Earlier this month, team owners argued it was their right to institute the lockout and suggested Nelson didn't have jurisdiction while the National Labor Relations Board considers an unfair labor charge filed by the league that players didn't negotiate in good faith.
Nelson disagreed, and said the NLRB proceeding shouldn't be used to affect the court case here.
The Associated Press contributed to this report.