Stalkers would only appear after Ezio has gained a considerable amount of notoriety, as he had proven himself to be a threat, and when approaching their target, they would blend in with the crowd and conceal their weapon behind their back, waiting for the right moment to strike. Should Ezio look towards them, stalkers would only stand still, appear to look away, and wait for an opportunity to attack him from behind.
They never engaged in open combat, and would only sneak up behind Ezio, stab him, and then flee. However, most of the time, Ezio would wrestle free of their grip and force their hand so that they killed themselves with their own blade. Alternatively, if Ezio recognized the stalker, either by seeing them sneaking up to him or by using Eagle Sense, he could then approach and kill them.
Stalkers would also participate in the attacks on Assassin Dens, climbing onto rooftops to kill Assassin defenders from behind. However, as overseer, Ezio could either shoot them with his Hidden Gun or send his Assassins to attack the stalkers directly.
- The appearance of a stalker is indicated by a white knife on the Animus mini-map, along with an ambient whispering noise and a change in the background music, as they get close to Ezio.
- The second set of the Romanies' guild challenges consisted of killing or countering five stalkers before they stabbed Ezio.
- If Ezio turns around just before a stalker is close enough to stab him, Ezio will forcefully push the stalker backwards and send them flying onto the floor. However, this does not kill the stalker, and they will flee if Ezio did not finish them off.
- Killing a stalker reduces Templar Awareness by 25%.
- Ottoman guards will chase down stalkers if they witness them assaulting Ezio.
- Stalkers generally carried around more money than other enemies. When Ezio or a thief would loot one, around 1000 akçe could be gained.