Abstract During phagocytosis, macrophages rapidly internalize a substantial fraction of plasma membrane without a net loss of surface area, suggesting that membranes are targeted to the cell surface from intracellular sites. Nevertheless, a requirement for mobilization of specific membrane compartments has not been demonstrated. We used bone marrow-derived macrophages (BMM) from wild type and vamp3 null mice to evaluate directly the requirement for this v-SNARE in phagocytosis of zymosan, IgG-beads, complement-opsonized particles, or latex microspheres. Regardless of the phagocytic receptor engaged or particle load, BMM lacking vamp3 exhibited no phagocytic defects when assayed after 1 h at 37°C, and phagosome maturation was unimpaired as judged by acquisition of lamp-1. In contrast, at early time points (5–15 min), internalization of zymosan (but not other particles tested) was significantly slower in vamp3 null BMM. These data indicate that vamp3 modulates efficient uptake of zymosan, but is not absolutely required for phagocytosis in primary macrophages.