This work aimed to develop an anionic cellulose nanofiber (CNF) bio-adsorbent from date palm tree waste and to investigate its removal efficiency compared to cationic methylene blue dye from contaminated water. Date palm pulp was first prepared from date palm leaves through acid hydrolysis using H2SO4, followed by hydrolysis in a basic medium using KOH, in which the process completely removed the components of hemicellulose, lignin, and silica. To obtain anionic CNF, the resulting pulp was further treated with H2SO4, followed by centrifugation. Biogel formation of the CNF suspension was promoted by sonication, where its removal efficiency of methylene blue dye was studied as a function of dye concentration, temperature, contact time, and pH value. In this work, we investigated two isotherms, i.e., Langmuir and Freundlich. The Langmuir model's consistency with the experimental data suggests that the adsorption of methylene blue dye onto CNF is monolayer and surface-limited. The reported maximum removal efficiency of 5 mg/g at 60 °C indicates the optimal temperature for adsorption in this specific case. Additionally, a pseudo-second-order model and Elovich model were also utilized to obtain a better understanding of the adsorption mechanism, in which we found not just physical adsorption but also an indication of a chemical reaction occurring between methylene blue dye and CNF. According to the results, that pseudo-second-order model's consistency with the experimental data suggests that the adsorption of methylene blue (MB) onto CNF is rate-limiting step involving chemisorption between the two. The study reveals that CNF adsorbents derived from renewable natural waste sources such as date palm leaves can be effective in removing cationic contaminants such as methylene blue dye.