The Italian was delighted with the way in which his troops responded to the unavailability of their star forward, praising Gareth Bale for the "most important goal" of his career
The Portugal international missed the Clasico clash through injury but saw wingers Angel Di Maria and Gareth Bale both score as Los Blancos ran out 2-1 winners at Mestalla.
"We deserved the title," the Italian said. "We pressed high up the pitch and we took advantage on the counter. We played really well and all the players worked hard, so congratulations to them.
"Cristiano's absence motivated the players," he added. "It made them work even harder. They all did well. You have to play as a team to beat Barcelona and we did that.
"Barcelona tried but we knew how to play against them. We needed to be compact and we were.
"We will take strength from this for the other two competitions."
Madrid face Bayern Munich in the semi-finals of the Champions League later this month and are still in the race for La Liga, although Atletico lead the Primera Division by three points with five rounds remaining.
Barca, meanwhile, look likely to end the season empty-handed following this loss, European elimination last week and their defeat at Granada on Saturday which saw the Catalans lose ground in La Liga.
"I don't think their cycle is finished," Ancelotti said. "They played a good game and even though things haven't gone that well for them this season, they will fight until the end in La Liga and remain a dangerous rival."
The Italian preferred to praise his players as a group, but believes that Bale's winner was the most significant strike of the Welshman's career to date.
"He played a very good game and it's definitely his most important goal. But all of the players did well.
"It was a fantastic night and the fans from both teams made it an extraordinary spectacle."