The referendum was suspended on Monday by the 12-judge court following a one-hour emergency meeting, a spokeswoman said.
Regardless of the court's decision, the regional government in Catalonia had already said it would continue preparations for the non-binding referendum in the event it was blocked. Catalonian President Artur Mas signed a formal decree over the weekend calling for the vote on November 9, despite Madrid's objections.
Spain's Prime Minister Mariano Rajoy announced the government's appeal of the referendum on Monday, saying in a speech that such a vote was not "compatible with the Spanish constitution."
He added that the secession of Catalonia was out of the question.
"There is nothing and no one, no power nor institution, that can break this principle of sole sovereignty," Rajoy said.
Catalonia's potential independence has raised concerns about Spain's already ailing economy.