Mitochondrial Ca2+ transport is a critical process in cellular physiology. One pathway for export occurs via Na+/Ca2+exchange (mito-NCX) machinery. TMEM65 has recently been shown to be crucial for mito-NCX activity, but its precise mechanistic role remains unknown. To evaluate whether TMEM65 forms a complex with NCLX, a protein previously proposed to facilitate mitochondrial Na+/Ca2+ exchange transport, and to gain insights into TMEM65’s potential functions, we sought to characterize potential interactors of TMEM65, performing co-immunoprecipitation on TMEM65 containing a C-terminal ID4 tag.