The ad, created by The Bank, pushes William Hill's football-betting offering and first airs on TV today (14 September). It is similar to previously seen ads, but has been spiced up by the new soundtrack.
It will run around the start of the Champions League group stages on channels including ITV, Sky Sports and ESPN.
According to EMI Music, no Sex Pistols song has been used in an ad before. The song will also be used in radio ads as part of the campaign.
The band has been paid an undisclosed figure for the track, and the four current members (John Lydon, aka Johnny Rotten, Steve Jones, Paul Cook and Glen Matlock) have each been given a £500 bet to make for a charity of their choice.
Media planning and buying for the campaign is by the7stars.
Ian Cassie, executive creative director of The Bank, said: "It’s testament to the team at William Hill that they took, what could be considered for many, as an unconventional approach to feature the Sex Pistols against graceful football action."
"What we wanted to achieve was a direct response ad that stood out by mixing a blend of the classic and the contemporary – featuring an iconic band like the Sex Pistols demonstrates what an iconic brand William Hill has always been."
Last week it emerged the band had put its name to a new perfume, which is already available in France and launched in the US on Friday (10 September).
Pistols frontman John Lydon has fronted a series of ads for Country Life butter, in which he extols the virtues of the spread, proclaiming that he doesn’t just eat it because it's British, but because he likes it.
He said his fee for the campaign was to fund the reunion tour for his post-Sex Pistols band, Public Image Limited.
Lydon recently told The Guardian: "I showed what I can do with butter, right? 85% increase in sales. I’m very proud of them Country Life ads. They were funny and clever and classy.
"But no one’s come in for me with anything else. Like Red Stripe lager. I drink lakes of the stuff. Why haven’t I been signed up for something like that? Or Marlboro fags? It amazes me that people don’t get the opportunity of me. I sell."
The Sex Pistols reformed for the first time since their 1970s heyday with original bass player Glen Matlock, for the Filthy Lucre tour in 2002.