The study of dendritic cell involvement in complex phenomena that rely on multi-cellular interactions, such as immune homeostasis, stimulation, and tolerization, called for the investigation of dendritic cell functions within physiological context. To this end we have developed a conditional cell ablation strategy that is based on dendritic cell-restricted expression of a Diphtheria Toxin receptor (DTR) using the CD11c/Itgax promoter. Here, we provide basic protocols that describe the use of this prototypic dendritic cell ablation model and highlight pitfalls and strengths of the approach.