We have cloned the cDNA encoding I kappa B-beta, one of the two major I kappa B isoforms in mammalian cells. The recombinant I kappa B- beta protein interacts with equal affinity to p65 and c-Rel and does not exhibit a preference between these Rel proteins. Instead the primary difference between I kapp B-alpha and I kappa B-beta is in their response to different inducers of NF-kappa B activity. One class of inducers causes rapid but transient activation of NF-kappa B by primarily affecting I kappa B-alpha complexes, whereas another class of inducers causes persistent activation of NF-kapa B by affecting both I kappa B-alpha and I kappa B-beta complexes. Therefore, the overall activation of NF-kappa B consists of two overlapping phases, a transient phase mediated through I kappa B-alpha and a persistent phase mediated through I kappa B-beta.