Through an examination of a Midwestern professional comedy club, this article theorizes stand-up comedy as part of the broader affective-cultural assemblage that is nightlife entertainment. Using the Deleuzian notion of assemblage, this analysis builds on poststructural accounts of the dynamic and transient properties of culture, and the relationship between space, culture, and affect. As a specific affective-cultural assemblage, stand-up comedy operates as both constrainer and enabler of racial and heteronormative order through the bringing together of a variety of diverse logics and practices. I argue that stand-up comedy should no longer be thought of strictly in discursive or symbolic-interactionist terms that over-determine the roles of particular agents (i.e. comics). Instead, when described as emergent, transient, and fundamentally affective, stand-up comedy and other cultural forms can be seen for their openness and multiplicity, both contributing to racial and heteronormative order as well as upending it.