After the release of the archangel Lucifer from his Cage in Hell, hunters Sam and Dean Winchester begin seeking the legendary weapon known as the Colt as a possible way to kill Lucifer. The Winchesters eventually learn from Becky Rosen that Bela Talbot had given the Colt to a demon named Crowley after stealing it from them and not to Lilith as the Winchesters had previously believed, giving them a lead on the Colt's location.
After learning of Crowley, the Winchesters employ fallen angel Castiel who follows Crowley from a crossroads deal to his mansion which is protected by angel warding. With the location of Crowley revealed, the Winchesters plan a raid to steal the Colt from him.
In order to create a distraction, hunter Jo Harvelle dons a dress and approaches the gate to Crowley's mansion and calls for help over the intercom, claiming that her car has broken down. Two of Crowley's demon guards approach and attracted to Jo, try to forcefully take her with them. Jo fights back, incapacitating one guard before Sam kills both with the demon-killing knife. The Winchesters thank Jo for her help and set up for the next phase of the plan by sneaking into the mansion and painting a devil's trap beneath a rug to trap Crowley and then having Jo cut the power to the mansion.
Drawn by the power outage, Crowley makes his way towards the Winchesters before spotting their devil's trap. Crowley berates them due to the expensive nature of the rug before they are captured by two more demons. Crowley reveals the Colt to them which he is aware they are after before unexpectedly using the legendary gun to kill his own two demons. Crowley then tells the Winchesters they need to talk privately.
Leading the Winchesters to his study, Crowley reveals that he purposefully spread rumors of his having the Colt so that they would know about it. When Sam questions why, Crowley tells them that its because he wants them to use the Colt to kill Lucifer. Crowley goes on to explain that Lucifer is an angel famous for his hatred of humanity and Crowley doubts that Lucifer cares about demons, who are twisted human souls, any more than he does humans. Crowley believes that to Lucifer, demons are just servants and once he's done with humanity, he will wipe out demons next, making Crowley's aid of them a matter of survival. Sam agrees and Crowley hands him the Colt and tells the Winchesters that Lucifer will be in Carthage, Missouri. With this information, Sam tries to shoot Crowley in the head at point-blank range with the Colt, but it is unloaded. Crowley warns the Winchesters not to miss, tosses Dean more bullets for the Colt and teleports away to "all points nowhere."
Using the information Crowley gave them, the Winchesters, Ellen and Jo Harvelle and Castiel attempt to kill Lucifer in Carthage, Missouri where he is raising Death. However, Lucifer turns out to be one of five things the Colt can't kill and survives.
Unknown to the Winchesters, during this encounter, Crowley has his valet slip a magic coin into the Impala so that he can listen in on them and track their progress. Crowley later returns to aid the Winchesters in finding the Four Horsemen of the Apocalypse and claims to have not been aware that the Colt would fail.
- This is the Winchesters first encounter and team-up with Crowley who they would go on to both fight against and with over the course of the series.