In a meeting of two of the true giants of European football, the Italians will be desperate for a win that would keep them in second position in Group B.Antonio Conte's men have been held to 1-1 and 2-2 draws against Copenhagen and Galatasaray, respectively, and will enter the fixture off the back of a disappointing 4-2 loss at Fiorentina on Sunday.
First-half goals from Carlos Tevez and Paul Pogba looked to have the Bianconeri side cruising, but Fiorentina – who were inspired by a hat-trick from Giuseppe Rossi – scored four times in the last 25 minutes to inflict Juve's first defeat of the season.
Conte hopes his side can respond quickly and denied claims that they were focusing on Madrid in the build-up to their trip to Florence.
"We lost a game that we dominated for 70 minutes. We were home and dry and had more than one opportunity to score the third goal," the former Italy international said after the defeat.
"I don't think our focus was already on Real Madrid, otherwise we would have got our overall approach wrong."
Defender Stephan Lichtsteiner (thigh) and midfielder Simone Pepe (thigh) have been ruled out of the fixture.
And Juve's injury worries do not stop there, with strikers Mirko Vucinic and Fabio Quagliarella (both thigh) also set to miss out.
Nine-time winners Madrid have won both of their Champions League fixtures this season, thrashing Galatasaray 6-1 before easing past Copenhagen 4-0.
Cristiano Ronaldo has scored five goals in those two matches, making him the competition's leading scorer.
The Portuguese star – who was top scorer last season with 12 goals – has now scored 24 goals in his last 20 Champions League appearances. Ronaldo's 56 career strikes in the competition also see him fourth in the all-time scorers' list.
World-record signing Gareth Bale has made just four appearances for Madrid since joining from Tottenham but returned from a thigh injury in the club's 2-0 win over Malaga on Saturday.
He is expected to feature against Juve, while midfielder Xabi Alonso has returned to training following a broken foot.
The match will come too soon for Alonso but the news of his return will be a significant boost for theBlancos, who can move a step closer to qualification for the Round of 16 with another victory.