There was a question mark over Bale's availability for the first-leg clash at the Calderon but Real's world record signing is "100 percent" fit and has been called up along with every other member of the squad, Ancelotti told a news conference.
The Welshman has been on patchy form this term after an impressive debut season in Spain, when he scored in Real's Champions League final win over Atletico and netted a memorable winner to beat Barcelona in the King's Cup final.
Ancelotti said he was sure Bale, who has been whistled by some fans at the Bernabeu stadium, would perform in the final weeks of the campaign, with Real seeking a record-extending 11th European crown and closing in on leaders Barca in La Liga.
"I have the good fortune to be able to count on everyone for such an important game," Ancelotti said.
"Bale had a fantastic season last term and scored in both finals," the Italian added.
"We hope to play more finals over the coming months and he will be important, like last year.
"We have to use our individual quality and all our weapons to play a game with speed and pace in attack."
Real must end a woeful recent run against their city rivals to reach the last four of Europe's elite club competition.
They have met six times this term, with Atletico triumphing over two legs to lift the Spanish Super Cup, twice winning in La Liga, including a 4-0 drubbing at the Calderon In February, and knocking them out of the King's Cup, also over two legs.
"We come into this match in good shape," Ancelotti said.
"It's nothing like February. We're fresh and we can play with intensity and at a high tempo.
"Personality and courage are very important and this team has that. Tomorrow I'd like to see a team that has control over the play and that competes throughout the whole match."
(Reporting by Iain Rogers, editing by Ken Ferris)
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