Harden will be reevaluated in approximately 10 days.
The nine-time All-Star had missed two games due to hamstring tightness and only saw four minutes of action in his return Monday before the new injury forced him out.
The 31-year-old has been an instrumental part of the Nets' surge to the top of the Eastern Conference, averaging 25.4 points, 11.0 assists and 8.7 rebounds through 34 games since arriving from the Houston Rockets in January.
Though Harden was originally headed to Brooklyn to supplement the team's All-Star duo of Kevin Durant and Kyrie Irving, all three have missed action throughout the season and only started six games together. Harden has missed games previously with a left thigh contusion and a sore neck.
Without Harden, Irving will be tasked with carrying the offensive load, while Bruce Brown should see an increase in minutes.