The Blades had not won, or even scored a goal, at Millwall for four years before this game. But strikes from Andy Liddell and Derek Geary accounted for the South Londoners, whose promotion challenge has tailed off in recent weeks.
This was Millwall's third successive defeat and they are losing touch with the play-off zone. Sheffield United, by contrast, are growing in confidence.
Millwall were without inspirational player-manager Wise who was suffering from a persistent calf injury, but midfielder Jody Morris returned after a hamstring problem.
Winger Paul Ifill, who came on and scored against Crewe in midweek, also started and Scott Dobie partnered former Blades striker Barry Hayles up front.
The visitors were without striker Luke Beckett after his wife gave birth to twins on Friday, so former Lions striker Paul Shaw started.
Millwall started brightly and could have scored within ten seconds, when Hayles charged down a clearance and forced keeper Paddy Kenny into a smart save.
The Blades fought back and Irish midfielder Alan Quinn hit a fizzing half-volley that required an acrobatic save from Graham Stack.
Both teams started the second half with ten men after an altercation in the tunnel between Kenny and Lions captain Kevin Muscat.
With no substitute keeper on the bench Sheffield United were forced to put defender Phil Jagielka in goal.
He was soon picking the ball out of the back of the net after Millwall defender Marcus Phillips ghosted in at the far post to head in a Peter Sweeney free-kick.
Rather than letting their heads drop United struck back to seal victory.
Liddell, just minutes after coming off the bench bent in a 25-yard free-kick in the 77th minute to level the scores.
Then, six minutes from time, the impressive Geary fired home a stunning 20-yard half-volley to seal the points.