On Thursday, Massimiliano Allegri's men went down 2-1 at home as two goals from Mohamed Salah gave Fiorentina the advantage in the first leg of their semi-final.
It was the reigning Serie A champions' first loss at Juventus Stadium in any competition since April 2013 - a remarkable run of 47 games.
As Juve turn their attentions back to the league, Padoin is in confident mood ahead of the visit of Sassuolo.
"We need to bounce back from this defeat and I'm sure we'll do so at the first time of asking on Monday," he told the club's official website.
"We know we'll be up against a decent team and a very young one at that.
"They were the first side to bring an end to our run of six consecutive victories at the start of the season, but winning is the most important thing on Monday.
"We've done well to gain a nine-point lead over Roma but with 13 games to go and 39 points up for grabs, it means nothing as things stand, anything can still happen."
That lead could be cut to six points by the time Juve kick off, with Roma visiting Chievo on Sunday.
"I don't think this is a difficult time for us, we just need to step up the tempo a little and finish games off earlier," Juve striker Carlos Tevez told Sky Sport Italia.
"No matter what some people say, nine points isn't enough of an advantage over Roma to relax. We have to speed up and get the Scudetto under lock and key as soon as possible."
Although Juve's only league defeat of this season came back in late October, Allegri's side have drawn three of their last five Serie A outings, while Sassuolo have lost each of their last three games, conceding eight goals in the process.
Former Juve man Simone Zaza has eight Serie A goals in 19 appearances this season - including his effort in the 1-1 draw between the two sides in the reverse fixture back in October.
The visitors, meanwhile, will have to keep a watchful eye on Tevez, who has 21 goals in all competitions this campaign, including three in his last five outings.
Juve's major injury concern surrounds experienced midfielder Andrea Pirlo, who has not featured since the UEFA Champions League win over Borussia Dortmund after picking up a calf problem.
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