The aim of this work is to investigate the adsorption of an anionic dye, the Murexide (MX) present in aqueous solution, on activated carbon, derived from prickly pear seed cake biomass after bio-oil extraction. The obtained adsorbent used was characterized by Bohem titration, pH of point of zero charge (pHPZC), FTIR spectroscopy, Brunauer–Emmett–Teller surface area (SBET), and scanning electron microscopy (SEM). The different experimental parameters of the adsorption process, such as temperature, contact time, initial dye concentration, and adsorbent dose, were studied. For the optimization of the process, the effects of these parameters were investigated using the full factorial experimental design methodology. Design Expert 18.104.22.168 Trial software was used for generating the statistical experimental design and analysing the observed data. Langmuir and Freundlich’s adsorption models were employed to provide a description of the equilibrium isotherm. The adsorption process was found to obey Langmuir, which indicates that the Murexide had formed a monolayer onto activated carbon. Furthermore, according to the regression coefficients, it was observed that the kinetic adsorption data can fit better by the pseudo-second-order model compared to the first-order Lagergren’s model. The thermodynamic studies indicated that the adsorption of Murexide occurs in a spontaneous and exothermic process. The regeneration process of the exhausted adsorbent was studied to assess the economic and operational feasibility. According to the obtained findings, it is proposed that the activated carbon prepared from prickly pear seed cake retains a high potential for Murexide removal and is suitable for repetitive usage.