At some point in the past, Kraven killed Hector Ayala, the former White Tiger.
Kraven first arrived in New York and started to antagonize Ava Ayala from afar. First was during a training session with her group, in which he caused her to act chaotic and run rampant. However, Ava was not easily fooled and knew he was behind it. During her attacks on New York, her team chased after her. However, Kraven appeared before the entirety of the group sans Ava and another and defeated Nova and Iron Fist with a knockout dart. As Power Man moved in to attack him, Kraven easily took him out with his knockout gas.
He soon stole the Tiger Amulet, but was transformed into a humanoid tiger. Ava and Peter removed it and White Tiger almost finished Kraven, but Spider-Man stopped her by telling her his own experience with the burglar who killed his uncle.
In the episode "Return of the Sinister Six", Doctor Octopus and Lizard spring the imprisoned Sinister Six members from Ryker's Island and equip them with high-tech armor made from stolen OsCorp technology. Kraven is given a lion-themed armor with razor-sharp mechanical claws. Kraven fights Spider-Man and then White Tiger once again. He also assists in the group's gang assault on Iron Patriot. Eventually, Spider-Man was able to defeat Kraven along with the other Sinister Six members.
In the episode "The Savage Spider-Man", Kraven teams up with Taskmaster in a plan to lure Ka-Zar to New York by kidnapping Zabu so that Taskmaster can recruit Ka-Zar to his side. During Taskmaster and Kraven's fight with Spider-Man, Ka-Zar and Wolverine, Kraven uses a dart with a special poison in it on Spider-Man which transforms him into the Man-Spider. Upon being pursued to Manhattan by Spider-Man, Wolverine and Ka-Zar, Kraven seeks to perform a ritual to become immortal by sacrificing Zabu at sunset (with the ritual occurring once every thousand years) while Taskmaster plans to get Ka-Zar on his side. With help from Wolverine who deals with Taskmaster, Spider-Man and Ka-Zar defeat Kraven who tries to knock Zabu off the bridge only for Spider-Man and Ka-Zar to catch Zabu before he can fall into the river. Kraven is later arrested by S.H.I.E.L.D.
In "Contest of Champions" Pt. 1, Grandmaster pairs up Kraven the Hunter with Molten Man and Wendigo King in the game of "Last Man Standing" against Collector's team of Spider-Man, Iron Man, and Hulk during. Kraven the Hunter coordinated Molten Man and Wendigo in the battle while using some of his traps. When chased by Hulk, Kraven used a special pollen that knocked out Hulk removing him from the game. After Spider-Man did tricks that removed Wendigo King and Molten Man from the game, Spider-Man engaged Kraven the Hunter and eventually defeating him allowing Collector to score the first victory against the Grandmaster.
In the episode "HYDRA Attacks" Pt. 2, Doctor Octopus frees Kraven the Hunter, Batroc the Leaper, and Boomerang from the brig of the S.H.I.E.L.D. Tricarrier-turned-HYDRA Island where he holds a contest where whoever can successfully capture Spider-Man will join his latest incarnation of the Sinister Six. Kraven the Hunter follows Spider-Man and Scarlet Spider into the sewers where they managed to bring some pipes down onto him. Later on, Kraven the Hunter confronts Spider-Man on HYDRA Island in an attempt to defeat him only to be knocked out of HYDRA Island by Scarlet Spider.
In the episode "Double Agent Venom," Kraven the Hunter sports new HYDRA armor as he hunts Spider-Man and Scarlet Spider. After abducting Scarlet Spider, Kraven the Hunter takes Scarlet Spider into space where Spider-Man discovers that they are heading to the returned HYDRA Island. After Spider-Man and Scarlet Spider free Flash Thompson following his separation from the Venom symbiote, Doctor Octopus throws Kraven the Hunter into the Venom symbiote which gives him the appearance of a humanoid lion. Flash was able to get the Venom symbiote off Kraven the Hunter and help defeat him.
In the episode "The New Sinister 6" Pt. 2, Kraven the Hunter appears as a member of Doctor Octopus' Sinister Six when they attack the Triskelion. During the battle, Kraven the Hunter uses a device that causes Cloak to lose control of his teleportation.