AbstractGraphene oxide (GO) is an excellent bacteria‐killing nanomaterial. In this work, macroscopic applications of this promising nanomaterial by fixing GO sheets onto cotton fabrics, which possess strong antibacterial property and great laundering durability, are reported. The GO‐based antibacterial cotton fabrics are prepared in three ways: direct adsorption, radiation‐induced crosslinking, and chemical crosslinking. Antibacterial tests show that all these GO‐containing fabrics possess strong antibacterial property and could inactivate 98% of bacteria. Most significantly, these fabrics can still kill >90% bacteria even after being washed for 100 times. Also importantly, animal tests show that GO‐modified cotton fabrics cause no irritation to rabbit skin. Hence, it is believed that these flexible, foldable, and re‐usable GO‐based antibacterial cotton fabrics have high promise as a type of new nano‐engineered antibacterial materials for a wide range of applications.