Second-placed Real, who hammered Granada 9-1 on Sunday with Portugal forward Cristiano Ronaldo netting his first five-goal haul, are four points behind Barca, who host Almeria earlier on Wednesday, with nine games left.
Barring last-minute injuries, Ancelotti will have both Isco and James Rodriguez available for the first time in two months since James broke a bone in his foot and must decide which of the two playmakers joins fixtures Toni Kroos and Luka Modric across the centre.
Isco, a Spain international who was suspended for the Granada game, performed well first during Modric's lengthy injury layoff and then during the absence of James and has become a huge favourite with the Real fans.
Ancelotti, by contrast, has been whistled by some sections of the Bernabeu crowd as Real stumbled since the turn of the year and if he drops Isco and the team stutter again the Italian could be in for a torrid time.
James returned to action against Granada and Ancelotti praised him after the game, suggesting he favours the Colombian, who joined from Monaco for a fee of 80 million euros (58.97 million pounds) in the close season.
"He played very well, it didn't look as though he hadn't played for two months," Ancelotti told a news conference.
"He showed his quality, his intensity and his ability to keep possession. He is a very direct player and always shows great vision and awareness."
After edging Celta Vigo 1-0 on Sunday, Barca will be looking for a more convincing performance at home to struggling Almeria, who sacked coach Juan Ignacio Martinez after Saturday's 4-1 home defeat to Levante.
Barca's Argentina forward Lionel Messi, in particular, will be keen to get back among the goals after failing to score in his last three outings in all competitions.
Ronaldo leapfrogged Messi to the top of the La Liga scoring chart with Sunday's five and has 36, with Messi, who holds the record for a single season of 50, on 32.
(Editing by Amlan Chakraborty)
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