The Spaniard, best known for his roles in Pain and Glory, The Mask of Zorro and as the voice of Puss in Boots, will star alongside Hollywood legend Harrison Ford, Phoebe Waller-Bridge, Mads Mikkelsen, Thomas Kretschmann, Shaunette Renee Wilson and Toby Jones.
Banderas said: “It is an [honour to] join on the screen… one of the most charismatic heroes in the history of cinema, with Harrison Ford, Steven Spielberg, James Mangold and the rest of this fabulous Indiana Jones team.”
Mangold has taken over directing reigns from Spielberg and has been seen in Glasgow on set.
Filming is currently under way in the city centre, with several streets transformed into the United States for the fifth movie in the franchise with 79-year-old Ford reprising the titular role.
Ford has yet to be spotted on set but a body-double has been seen riding on horseback past a parade scene.
The veteran actor injured his shoulder in a fight scene rehearsal ahead of filming on location in Scotland last month.
One of the floats in the parade featured the words “Apollo 11” which, along with other references to the astronauts, suggests at least some of the film could be set in 1969.