The Iron suffered a 2-1 defeat at the hands of Gillingham, on a frustrating night at the MEMS Priestfield Stadium.
Following on from Saturday's 2-1 success over Shrewsbury, Mark Robins opted to leave things as they were in terms of team selection.
Luke Daniels started in goal, while in front of him there was a back four of Scott Wiseman, David Mirfin, Murray Wallace and Scott Laird. Neal Bishop started once more alongside stand-in captain Jack King, with Gary McSheffrey, Tommy Rowe and Paddy Madden providing the support to Tom Hopper up-front.
It was a tough start for the Iron, as the hosts immediately applied pressure. However, the side which started the night at the summit of League One, failed to create any clear cut chances early on.
It was in fact United who almost edged in-front, when McSheffrey went close in the 15th minute. The winger picked the ball up on the right-hand side, before unleashing a curling left-footed effort, which flew inches wide of Gills 'keeper Stuart Nelson's post.
The Iron number 28 threatened again, five minutes later. This time he made Nelson work, with a free-kick from the edge of the penalty area. But, despite hitting the target, it was comfortable work for the hosts' stopper.
McSheffrey nearly turned provider half-way through the opening period, when Wallace glanced a header fractionally over the crossbar. The loanee stole a yard at the near post, after McSheffrey had superbly delivered a corner from the right-hand side.
Robins' side were in the ascendancy and continued to make inroads. Ten minutes before the break, Laird precisely slipped Rowe in, and after jinking past two Gills defenders in the box, he was ultimately foiled as he shot for goal.
There was a real crispness to United's play and everyone was keen to get involved, including full-back Wiseman, who poked wide after beating West Bromwich Albion loanee Bradley Garmston on the right touchline.
The quality of McSheffrey's delivery was causing the home side real problems. Shortly before the interval, he whipped another brilliantly flighted corner into the near post, which Nelson flapped at and almost turned into his own net.
United did everything but break the deadlock and would have been frustrated to have gone in at half-time level.
The Iron's dominance was finally rewarded, eight minutes into the second half. Wiseman floated a cross from the right flank, which evaded almost everyone. One man it didn't miss though was Laird, who arrived late into the penalty area and struck a beautiful left-footed volley, which flew past Nelson and into the roof of the net.
The goal was no more than the visitors deserved, after dominating the vast majority of the first period and the opening stages of the second.
However, despite getting their noses in-front, United were soon pegged back – as, five minutes later, substitute Brennan Dickinson powered home a header from close range. The forward wasn't picked up from a corner and had the simple task of heading home from six yards out.
Dickenson, who brought the sides level, almost gave his side the lead moments later with a long range effort. The attacker found space in the middle of the park and hit a volley which looked to have beaten Daniels. Fortunately enough though, the ball flashed just wide of the Iron 'keeper's left hand post.
There was a great sight for everyone connected with the club shortly after, as Luke Williams made his return to first team action, as he replaced the dangerous McSheffrey. Williams had been out of action since the opening day of the season, but was given a standing ovation by the traveling Iron support.
The visitors were inches away from re-taking the lead with 15 minutes to go, after Laird drove a dangerous ball across the six-yard box. There were three Iron players queuing up at the far post, but none of them could quite get there in time and the ball drifted out for a goal kick.
The game's flashpoint came with ten minutes remaining, as former United midfielder Josh Wright went in late on Madden and much to the hosts' surprise, Wright was dismissed.
This then gave the Iron an added impetus to go on and win the game and they almost did when King curled an effort towards the top right corner of the goal. However, the midfielder got a little too much elevation on his attempt.
There was to be one final twist however, as Harry Lennon won it late on for Gillingham. Much like Dickenson's equaliser, the Gills man wasn't picked up and had the simple task of finishing from close range.
Wallace almost dramatically brought United back level with seconds to go, but was denied from point blank range by Nelson.
Shortly after, referee Lee Swabey brought an end to proceedings and an end to a run of three successive victories for the Iron.
Gillingham: Nelson, Garmston, Egan, Hessenthaler (McGlashan 46), Williamson (Dickenson 46), Morris, Osadebe (Houghton 71), Donnelly, Dack, Lennon, Wright.
Substitutes not used: Morris, Ehmer, Oldaker, Mbo.
Scunthorpe United: Daniels, Wiseman, Mirfin, Wallace, Laird, Bishop, King, McSheffrey (Williams 66), Rowe, Madden (Lolley 90), Hopper (van Veen 84).
Substitutes not used: Anyon, McAllister, Goode, Clarke.