Isco will be sidelined for Real Madrid's first home game of the La Liga season against Celta Vigo on Saturday, live on Sky Sports 3 HD and Sky Sports Mix from 7.10pm.
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The 24-year-old midfielder has picked up a knock on his ankle and will not feature in the game at the Bernabeu, while manager Zinedine Zidane reiterated the importance of all members of his squad.
Karim Benzema and Cristiano Ronaldo - who won the UEFA's Best Player in Europe award for the 2015/16 season on Thursday - also remain unavailable, while rumours continue to swirl about the future of James Rodriguez and Raphael Varane.
"He's got some discomfort in his ankle. Last week he took a knock on his ankle and he's still not recovered. He left the training session and he won't be with us tomorrow," he told a pre-match press conference.
"Anything can happen until the 31st. Nobody is coming in, that's certain, but it's possible some may leave. This is the squad I've got and I'm happy with the players I have.
"There are injured players and some who still haven't come back yet. There's good depth, the season will be tough and we're going to need all of our players.
"I have 23 or 24 players and they're all going to get a game. Cristiano and Karim aren't playing and there are other players who have done well. When everyone is back, I'll make the decisions about who will play, that's not going to change.
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"I want to be clear, every player is important and it's no surprise that they're all here. If they're playing for Real Madrid, it's because they're very good footballers."
Gareth Bale made a goalscoring return to the squad in their opening La Liga fixture of the season - netting twice as Los Blancos beat Real Sociedad 3-0 last weekend - and Zidane is looking forward to being back on home turf.
"It's exciting, it's the first home game and we want to play as well as we can," the former France international added.
"We started very well in La Liga at a difficult ground. Now we're up against a very good opponent, we know all about their quality and we're ready to play tomorrow. The fans will come to the ground and it's great for us to see a full stadium.
"We've got another tough opponent who play some good football, but our plan is the same as ever. It's our first game at home in front of our fans and we've got to come out the blocks well to try and get the three points."
Celta Vigo lost their opening La Liga fixture to new boys Leganes at the Balaidos Stadium, with Marcelo Diaz, Claudio Beauvue and goalkeeper Ruben Blanco Veiga injured for the trip to the Bernabeu.