The club said Friday it was filing an appeal with the country’s sports tribunal to annul the suspension that was keeping Neymar from playing at the Santiago Bernabeu on Sunday.
Barcelona had already filed an appeal with the Spanish football federation, but its demand was rejected on Thursday.
Neymar was handed a two-game suspension for sarcastically applauding an assistant referee after being sent off in Barcelona’s 2-0 loss at Malaga on April 8.
The two-match ban was in addition to the one-game suspension he was automatically required to serve for the red card. He already served that match against Real Sociedad last weekend.
Two-time defending champion Barcelona trails Real Madrid by three points with six rounds to go in the Spanish league. Madrid, seeking its first league title since 2012, also has a game in hand.