Cellulose nanofibers were prepared using TEMPO/NaBr/NaClO oxidation of kraft pulp and successive ultrasonic treatment, and the properties were characterized by conductimetric titration, X-ray diffraction, and atomic force microscopy. The resulting product was then applied as an anionic microparticle to constitute a microparticulate system with cationic polyacrylamide (CPAM), to induce the flocculation of the kaolin clay suspension. The flocculation effect was evaluated by determining the relative turbidity of clay suspension. The results showed that the obtained cellulose nanofibers had cellulose I structure with higher crystallinity than that of the kraft pulp, and their cross-sectional dimension was in the range of 3–5 nm. They had more negative zeta potential at neutral and alkaline conditions. It was found that the microparticulate system showed high flocculation effect on kaolin clay at a very low level of nanofiber addition, and a high shear level after CPAM addition was helpful for the flocculation. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40450.