Coaches will be able to make their one substitute per game, which replaces the fourth member of the interchange bench, at any time during play.
Clubs will need to officially name their substitute - who will wear a green vest while on the bench - when the final teams are lodged 90 minutes before the bounce.
"It's not required that there's an injury; there will be three interchange players as per normal and one [other player] will be the substitute," AFL general manager of operations Adrian Anderson said on Monday.
"When the substitute comes onto the ground, the player that comes off - the substituted player - can no longer take any part in the game."
The teams, spectators and media will be informed of an activated substitute via the scoreboard and the player coming off will don a red vest.
Penalties will apply to teams that don’t follow the procedures that were outlined to clubs on Monday morning.
"It depends on the nature of the error but if for example like with the normal interchange there are too many players on the ground or they haven't notified the interchange steward, then a free kick and a 50m penalty can apply," Anderson said.
"If it’s a procedural error, then financial sanctions can apply."
The rule was announced at the start of October after the Laws Committee decided to tackle growing concerns surrounding congestion, fairness and increasing injuries.
Anderson said he expected the clubs to positively accept the details of how the rule would be enforced.
"The main news has already arrived in terms of the guts of what's going to happen and how it's going to work," he said.
"This is really about the details of operationally how it's going to work on match day so I'm sure the clubs will work through that in the professional manner they usually do."
The Toyota AFL Premiership Season gets under on Thursday March 24 with Carlton versus Richmond at the MCG.