The Sec23p/Sec24p complex is a component of yeast coat protein II (COPII), the coat protein complex responsible for vesicle budding from the endoplasmic reticulum (ER). Database searches and molecular cloning reveal that four different mammalian Sec24p-like proteins exist, all with about 20% amino acid identity with the yeast Sec24p. Sec24A and Sec24B share about 50% amino acid identity. Sec24D is cloned by screening a human pancreas of cDNA library with an expressed sequence tag (EST) fragment that is homologous to, but distinct from, Sec24A and Sec24B. Sec24D shares about 50% amino acid identity with the gene product of KIAA0079, which we have designated as Sec24C. These mammalian Sec24s appear to form two subclasses based on homology. Sec24A/B and Sec24C/D share about 20% identity with each other and with the yeast Sec24p. The Sec24 sequences also share weak but significant homology to the mammalian Sec23A and Sec23B. Northern blot analysis revealed that Sec24C is ubiquitously expressed. Although Sec24D transcripts are detectable in all tissues examined, they are selectively enriched in certain tissues, particularly placenta and pancreas. myc-tagged Sec24C and sec24D colocalized with Sec13, another COPII component. This colocalization suggests that Sec24C and Sec24D are indeed associated with COPII structures on membranes of the ER–Golgi boundary. The existence of at least four forms of Sec24 in mammalian cells suggest that multiple forms of COPII complex may be involved in ER export.