In a statement released late on Sunday, UGT and CCOO, Spain's biggest unions, the Catalan National Assembly (ANC), a powerful pro-independence civil association, and 41 other organisations called for the work stoppage due to "the grave violation of rights and freedoms."
"We call all society, on employers' organisations, business owners, unions, workers, self-employed workers, institutions and all the citizens of Catalonia to stop the 'country' on Tuesday, October 3," they wrote.
Pro-independence Catalans often refer to the northeastern region as a "country."
The regional government says more than 840 people received medical attention on Sunday after riot police moved in on polling stations as Catalans tried to vote in the referendum, defying a ban by Madrid.
In some cases, police used batons and rubber bullets to evict people from the voting centres and confiscate the ballot boxes.
Spain's interior ministry said 33 police agents also received medical aid.
Madrid had warned Catalan separatist leaders they could not hold the vote in a region deeply divided over independence, stating it was illegal and courts had ruled it unconstitutional.
But they had retorted that Catalans had a right to decide on their future and pressed ahead anyway.
Catalonia is a major engine for Spanish growth, and accounts for around 19 percent of its GDP.
As such, any region-wide strike will be damaging for Spain.