Protocol for the detection of RHIM-protein oligomeric complex formation using DSP-mediated crosslinking in HEK293T cultured cells

Proteins harboring the RIP homotypic interaction motifs (RHIMs) can assemble into higher-order structures through RHIM-RHIM interactions. Here, we present a protocol for detecting RHIM protein oligomerization, which is central to cell death and inflammatory signaling. We describe the steps for expressing RHIM proteins in HEK293T cells, crosslinking their oligomers, and detecting them by immunoblotting analysis. This protocol can facilitate the study of RHIM-based oligomerization and its role in modulating cell death and inflammation. For complete details on the use and execution of this protocol, please refer to Mishra et al.¹.