The singer tweeted on Wednesday: "Join me & Target tonight for just one song. Let's 'Take Back the Night.' First 150 get in. See you there!"
Fans jammed the street outside Maxwell's in Hoboken, and police used loudspeakers to control the crowd.
The Jersey Journal reports that Timberlake's wife, Jessica Biel, was spotted near his trailer.
The indie-rock club closed its doors after a 35-year run in July.