The standard edition of the album contains 17 tracks; the "special edition" contains a second CD with 13 additional tracks. EMI also released a DVD compilation with 21 music videos, including 9 never released on DVD before. The compilation debuted at number 4 in the UK album charts, and critical reception to the compilation was generally positive.
The compilation contains only songs to which EMI holds the publishing rights, and so contains no songs from Radiohead's seventh studio album, In Rainbows (2007), released the previous year and recorded and released without a record label. Radiohead stated in 2003 that they intended to never release a "greatest hits" album. Radiohead: The Best Of was prepared when Radiohead decided not to renew their contract with EMI. In 2008, singer Thom Yorke said,:"There’s nothing we can do about it. The work is really public property now anyway... It’s a wasted opportunity in that if we’d been behind it, and we wanted to do it, then it might have been good."
The version of disc one for the US contains a bonus track, "Optimistic" (Edit), before "Fake Plastic Trees". The reasoning for its inclusion was that "Optimistic" was played on US alternative radio to promote Kid A—an album without any official singles—and proceeded to peak in the top 10 of the Billboard Modern Rock Tracks chart, one of only three Radiohead songs to do so.