The Barcelona man has struggled to reach full fitness since sustaining the ankle problem in a goalless draw against Athletic Bilbao on March 3.
De Jong has played just three matches since being forced off the at San Mames, appearing in both Champions League quarter-final matches against Paris Saint-Germain as well as April's Clasico.
However, De Jong has not kicked a ball in anger since being withdrawn in first-half stoppage time against Real Madrid.
Netherlands boss Ronald Koeman still decided to include De Jong in his Euro 2024 squad despite the obvious concerns over his fitness.
De Jong to miss Euro 2024 through injury
Unfortunately for the 27-year-old, he has ultimately lost the race to recover in time for this summer's tournament in Germany.
In a post on X
De Jong took to Instagram to comment on his absence, stating: "I'm sad and disappointed that I won't make it to the European Championship. We've been doing a lot of it the past few weeks, but my ankle needs more time unfortunately.
"It's a dream and greatest honour to represent our country in a final tournament. Wearing the orange shirt, singing the Wilhelmus and feeling the support of the entire country. But now I will, like the entire orange legion, cheer for our team from the sidelines. Let's go, boys."
De Jong's absence represents a considerable blow to the Netherlands due to his technical ability and wealth of international experience.
The Barcelona midfielder has earned 54 caps for his country and started each of his country's matches at Euro 2020 and the 2022 World Cup.
What are Koeman's alternative midfield options
Although De Jong's injury represents a significant setback, Koeman can still call upon a number of top-quality options, including Jerdy Schouten and Joey Veerman.
The PSV Eindhoven duo are among the contenders to feature in the Netherlands's double-pivot, while Koeman has the option of turning to Ryan Gravenberch and Georginio Wijnaldum.
Atalanta's Teun Koopmeiners is another midfield option, but there are concerns over his fitness after he was forced to withdraw from Monday's 4-0 win over Iceland.
The 26-year-old complained of some muscle discomfort in the warm-up and will now have to be assessed ahead of his country's tournament opener against Poland on June 16.
The Oranje will complete the group stage will face 2022 World Cup finalists France in their second game, before they wrap up the group stage with a game against Austria.