In Drosophila, the IMD pathway is indispensable for proper innate immune responses. Infection by gram‐negative bacteria elicits a signaling cascade culminating in the rapid induction of antimicrobial peptide gene expression by the NF‐κB transcription factor Relish. Signal‐dependent activation of Relish is an essential component in the IMD pathway and is regulated by the Drosophila melanogaster IκB Kinase (DmIKK) complex. DmIKK is composed of two subunits: the catalytic subunit IKKβ and non‐catalytic subunit IKKγ. Like many protein kinases, DmIKK must be activated for proper catalysis. Although there has been extensive research into the signaling pathways of the Drosophila innate immune response, little is known about the molecular mechanisms that lead to DmIKK activation. Here we report expression, purification, and in vitro characterization of a catalytically active multi‐subunit DmIKK complex. Support or Funding Information This work is supported in part by the California Metabolic Research Foundation. S. Cohen is a recipient of the Arne N. Wick predoctoral fellowship.