Nightin' Ale's is a fictional bar and strip club in the Venture Bros television show. It sometimes frequented by Brock Samson and Dr. Venture. Hunter Gathers also formerly worked there as an exotic dancer after his gender reassignment surgery, though he also used it as a base for his intelligence operations at the same time. The unnamed bartender at Nightin' Ale's from time to time engages in creative insults of both customer and employee alike. He is not seen in the Season 4 episodes that revisit Nightin' Ale's after it has been given a redesign by the shows' creators.
In Mid-Life Chrysalis after Brock has lost his OSI license, Dr. Venture and Brock Samson visit the club and orders some drinks. As the bartender serves them, he insults Brock. Brock is infuriated but restrains himself, as his license to kill has been removed. Although Dr. Venture flounders gaining the attention of the bar's dancers, Brock Samson enters the bathroom meets a blue-haired dancer in the bathroom for a sexual encounter. Memories of his OSI days and his now inability to legally kill ruins the mood for him, so Brock breaks it off. Meanwhile, Dr. Venture seemingly hooks up with a woman named Charlene. However, this is secretly part of a sinister plot by the Monarch that is resolved later in the episode. After Brock regains his license at the end of Mid-Life Chrysalis, he returns to Nightin' Ale's and gives the bartender a good thrashing.
In ORB and The Family That Slays Together, Stays Together (Part II), Brock Samson returns to the bar to gain information from Hunter Gathers regarding the Orb and later the OSI assassins sent to kill him. At this time, Hunter Gathers is working as one of the exotic dancers but takes a break to speak with Brock under the guise of speaking to a client. As Hunter takes too long, the bartender heckles her to get back to work.
Following Perchance to Dean, Nightin' Ale's receives a spruce up, and the general atmosphere seems livelier. In Self-Medication, Action Johnny, Dr. Venture, and several other former boy adventurers investigate the murder of their psychiatrist and find a matchbox that leads them to Nightin' Ale's. Their they find a supposed henchman of Dr. Z and move to question him. However, a biker named Francis cuts in, reminding Action Johnny that he has been banned and telling them to leave. Wonder Boy II steps in, says that if Francis messes with one boy adventurer, he messes with all of them. Action Johnny asks Francis if the herpes on his lips came from kissing his wife's ass after Johnny gave it to her. A huge brawl ensues, leaving the former boy adventurers wounded but victorious in the fight. However, their lead regarding the henchman is blown. Therefore, they leave the bar and go directly to Dr. Z' residence.