For the first time since 2010, Ronaldo will not be in the Champions League semi-finals.
And he held his head in his hands in despair as he slumped to the pitch at the Allianz Stadium.
He had single-handedly lifted Juventus into the quarter-final with a stunning hat-trick in the second leg against Atletico Madrid.
But he was unable to lift his team to a Champions League title he had won in the three previous seasons and in 2014 with Real Madrid, as well as with Manchester United 11 years ago.
He holds the record for goals scored — which includes 65 goals in the knockout rounds, another record.
Rival Lionel Messi is second on 110 goals after his double for Barcelona against Manchester United on Tuesday.
“Ajax deserved to qualify and you saw that on the pitch,” said Juventus president Andrea Agnelli, who said Allegri would be continuing as coach.
Ironically Agnelli — who is also president of the European Clubs Association (ECA) — is the driving force behind moves to change the Champions League and make it even harder for teams from smaller countries, like Ajax, to qualify.
Agnelli, 43, stressed that the team was growing.
“For our part, we are aware of having been in the Champions League quarter-finals for five or six years and are proud to continue this path,” said Agnelli.
Although Juventus are on course to seal an eighth consecutive Serie A title next weekend, they are out of the Coppa Italia, which they had won the four previous seasons.
“I’m looking at the big picture,” Agnelli said.
“We played a great return match against Atletico, and just a few years ago Juventus were 43rd in the European rankings, today we are fifth and this is the sign of the progress that continues.
“A few years ago, the Champions League was a dream, this summer it was a goal and it will be next year too.”
Ronaldo was a heavy investment with the club announcing expected losses for this season, having gone 19.2 million euros into the red for the 2017/2018 campaign.
Ajax, meanwhile, reached their first semi-final since 1997.
“You could see that Juve was a little scared of us and so were Real Madrid,” said coach Erik ten Hag. “It is very difficult to play against our style of football, as we have very technically-gifted players.”