Mr Trump’s Twitter account shared the video, which consists of a montage of photos from some of his past rallies, on Wednesday.
“We Will Rock You”, Queen’s hit dating back to 1977, plays in full during the two-minute clip.
A representative for the band told The Independent that “Queen did not agree to the use of the song” and that ”the band’s music publishers are removing it”.
A representative told BuzzFeed reporter Adam B Vary, who tweeted about the issue, that Queen “has already entered into a process to call for non-use of Queen song copyrights by the Trump campaign”.
The Independent has contacted the White House for comment.
This isn’t the first time Queen has opposed the use of one of the band’s tracks by the US president.
In July 2016, the band said that Mr Trump’s use of “We Are The Champions” at the Republican National Convention was “unauthorised” and “against our wishes”.
Brian May later said that even if Mr Trump had asked to use the song, his request would have been denied, calling Mr Trump a “very dangerous man” on Good Morning Britain.
While candidates may be able to use various songs during rallies, depending on the event’s specific conditions, the situation is different when it comes to using a tune in a campaign ad.
According to guidelines issued by the American Society of Composers, Authors and Publishers, “this kind of use may involve rights such as synchronisation of music with video and the possible use of the master sound recording”.
“The campaign will need to contact the song’s publisher and possibly the artist’s record label to negotiate the appropriate licenses with them. And remember, campaign videos containing music that are posted on the Internet also require these licenses,” the guidelines continue.
“Once the commercial has been produced, the TV and radio stations, and any websites that transmit the commercial, must have a public performance license.”