The 60-year-old Queen of Pop kicked off her headline set in Tel Aviv, Israel, by turning the stage into a makeshift chapel with a hooded choir and religious imagery as she revived the 1989 hit which turns 30 this year.
Madonna, who was dressed in a lengthy robe and dazzling diamond encrusted eyepatch, slowly walked down a steep set of stairs as she belted out the words to one of her biggest his.
She then switched up the atmosphere, giving her live debut of her new reggae-tinged track 'Future' featuring Migos star Quavo - who was proud to perform with "one of the best" pop stars in the world - and the pair brought their energetic performance to a dramatic close as the stage went black and they fell in unison backwards off a high platform.
Before hitting the stage, Madonna gave an impassioned speech reminding people to "never underestimate the power of music to bring people together".
She said: "Another really obvious statement is look at all the delegates behind us everybody here is from all over the world, right.
"Some of the counties that I have been privileged enough to visit and experience and the one thing that brings me to those countries and the thing that brings all of these people here tonight is music.
"So let's never underestimate the power of music to bring people together.
"To quote an amazing song [Madonna's 2000 hit 'Music'], 'Music makes the people come together.'"
She added: "I'm bringing the heat tonight!"
Asked what advice she would give the entrants, she said: "The first thing I'd like to say is that you are all winners no matter what happens.
"The reason that I believe that is because to get here where you are right now is not easy, right.
"First you had a dream and you had to believe in that dream and make many sacrifices and give a lot of things up.
"Earn your way to the place you are right now and in my opinion that makes a winner no matter what."
Madonna