Previous studies have suggested that neutrophils are required for resistance during infection with multiple pathogenic microorganisms. However, the depleting antibody used in those studies binds to both Ly6G and Ly6C (anti‐Gr‐1; clone RB6‐8C5). This antibody has been shown to deplete not only neutrophils but also monocytes and a subset of CD8⁺ T cells. Recently, an antibody against Ly6G, which specifically depletes neutrophils, was characterized. In the present study, neutrophils are depleted using the antibody against Ly6G during infection with the intracellular bacterium Listeria monocytogenes (LM). Our data show that neutrophil‐depleted mice are much less susceptible to infection than mice depleted with anti‐Gr‐1. Although neutrophils are required for clearance of LM, their importance is more pronounced in the liver and during a high‐dose bacterial challenge. Furthermore, we demonstrate that the protection mediated by neutrophils is due to the production of TNF‐α, but not IFN‐γ. Additionally, neutrophils are not required for the recruitment of monocytes or the generation of adaptive T‐cell responses during LM infection. This study highlights the importance of neutrophils during LM infection, and indicate that depletion of neutrophils is less detrimental to the host than depletion of all Gr‐1‐expressing cell populations.