The dicamba and glyphosate (DpG) tank mixture has been widely adopted for broad-spectrum weed control. However, recent studies indicated antagonistic interactions between these herbicides. Furthermore, little is known about the effect of non-ammonium sulfate water conditioner (non-AMS WC) adjuvant on the activity of DpG tank mixture. Thus, the present study was elaborated to evaluate (1) the interaction of DpG tank mixtures combinations on barnyard grass, and glyphosate-susceptible (GS) and -resistant (GR) horseweed control, and (2) the effect of non-ammonium sulfate water conditioner (non-AMS WC) on the interaction of those two herbicides on the aforementioned weed species. Greenhouse experiments were conducted in 2020 at the Pesticide Application Laboratory in North Platte, NE. Herbicide treatments were arranged in a two-level factorial design of six dicamba rates by six glyphosate rates for Study 1, and in a three-level factorial design of two adjuvant treatments (presence or not of non-AMS WC) by four dicamba rates by four glyphosate rates for Study 2. Both trials were conducted as randomized complete block designs with four replications and two runs. Antagonistic interactions were observed throughout DpG treatments in GS and GR horseweed. For instance, dicamba (560 g ae ha−1) and glyphosate (1260 g ae ha−1) resulted in 72% of GR horseweed biomass reduction, compared to 81% of estimated biomass reduction. For barnyard grass, antagonistic interactions were only observed within the reduced glyphosate rates. The addition of non-AMS WC had no effect on DpG antagonism. However, it improved the barnyard-grass control of glyphosate.