We have investigated the effect on sensitization of altering the area of application of 2,4,dinitrochlorobenzene (DNCB) at a constant dose per unit area. We showed that, when an area of less than 1 cm2 is used, this area is critical in determining the degree of sensitization. This contrasts with previous work that showed, for larger areas, an alteration in the area of application had little effect on sensitization, whereas keeping the area constant and increasing the concentration of DNCB increased the degree of sensitization. We suggest that not only is the amount of antigen important in determining response, but also the distribution of the antigen as presented to the afferent limb of the immune system.