The footage came into the public domain two days after Gray was shown making controversial comments about assistant referee Sian Massey.
Gray had his contract terminated following the release of the new footage, 24 hours after he and fellow presenter Richard Keys were disciplined for their remarks about Massey. Sky Sports reporter Andy Burton was also disciplined on Tuesday afternoon after footage emerged of him and Gray commenting on Massey's appearance.
Announcing the departure of Gray, who has been one of the faces of the broadcaster's live Barclays Premier League coverage since it began almost 20 years ago, Sky Sports said in a statement: "Sky Sports terminated its contract with Andy Gray.
"The decision, which is effective immediately, was made in response to new evidence of unacceptable and offensive behaviour.
"The new evidence, relating to an off-air incident that took place in December 2010, came to light after Andy Gray had already been subjected to disciplinary action for his comments of 22nd January 2011."
The new footage, which was recorded last month but only came to light in the last 24 hours, appears to show Gray making a suggestive comment towards colleague Charlotte Jackson, who does not openly react while he and Keys then burst out laughing.
Gray's sacking was followed by an announcement from Professional Game Match Officials that they had withdrawn 25-year-old Massey from the npower League Two match between Crewe and Bradford, where she had been due to run the line.
PGMO general manager Mike Riley said: "PGMO and Sian believe that, with any football match, the focus should not be on the officials but on the players and the game itself. Sian is an excellent professional who has unwittingly found herself in the middle of a story that has nothing to do with her competence as a match official.
"Sian only wants to be notable for her performance as an assistant referee and is keen for things to get back to normal so she can return to officiating as soon as possible. Sian has the full backing of PGMO and we hope she continues her development, which has shown excellent progress so far."
Source: PA
Source: PA