C-type lectin receptors (CLR) classically bind specific types of glycan moieties displayed on pathogens or damaged cells. Some family members bind endogenous ligands and mediate alternate functions such as cell adhesion, migration, or LN expansion/contraction. We discovered CD302, the simplest type I CLR with a single C-type lectin like domain (CTLD), showing it to be mainly expressed on myeloid phagocytes in human blood. Unlike related but more complex CLR such as CD205 (DEC205) and CD206 (MMR), CD302 colocalised with podosomes and lamellopodia structures in human myeloid cells (DC), leading us to hypothesise a role for this molecule in cell adhesion or migration. In this study, we used murine models to gain further insights into CD302 expression and its immunological function. Mouse CD302 transcripts were, as in human, highest in the liver, followed by lungs, lymph nodes (LN), spleen and bone marrow. Development of a mouse monoclonal antibody revealed a low molecular weight CD302 isoform in liver, with expression in hepatocytes, liver sinusoidal endothelial cells and Kupffer cells. A higher molecular weight form was detected in immune cells. Detailed analysis of CD302 transcription in mouse immune cells revealed highest expression by macrophages, granulocytes and myeloid dendritic cells (mDC). Interestingly, 2.5-fold more CD302 was observed in LN migratory DC that traffick into LN compared to resident mDC populations. Higher expression of CD302 in mouse M1 versus M2 macrophages was also noteworthy. The first CD302 knockout (CD302KO) mouse strain was generated, which exhibited a decrease in frequency and numbers of migratory mDC within LN compared to wild-type LN. CD302KO and wild-type DC had an equivalent capacity to undergo maturation, prime T cells, uptake antigens, and migrate towards the CCL19/21 chemokines. However, CD302KO migratory DC exhibited reduced in vivo migration into LN, confirming a functional role for CD302 in DC migration.