When approaching their target, stalkers 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, they would only stand still, appear to look away and wait for an opportunity to attack him from behind.
Stalkers 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 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, sneaking 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.
- An appearance of a stalker was 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 got close to Ezio.
- The second set of the Romanies' Guild challenges consisted of killing or countering five stalkers before they stabbed Ezio.
- If Ezio turned around just before a stalker was close enough to stab him, Ezio would forcefully push the stalker backwards and send them flying onto the floor. However, this did not kill the stalker, and they would flee if Ezio did not finish them.
- Killing a stalker reduced Templar Awareness by 25%.
- Ottoman guards would chase down stalkers if they witnessed them assaulting Ezio.