The textile materials functionalized with nanostructures have proven to be useful for many applications, such as antimicrobial, ultraviolet (UV) light protection, and self-cleaning substrates. The objective of this research is to synthesize and characterize zinc oxide (ZnO) nanoparticles (NPs) for the applications of UV absorbers and antibacterial activities. ZnO NPs were synthesized at different temperatures and reaction media of water (S-1) and 1,2-ethanediol (S-2) using precipitation and in situ methods on the surface of cotton fabric. The average crystalline size of the ZnO NPs estimated from the Debye Scherrer formula was found to be 32 and 26 nm for S-1 and S-2, respectively. The morphology of ZnO NPs characterized by scanning electron microscope revealed that agglomerated nanostructures were homogeneously formed on the fabric surface for S-1 and S-2; on the other hand, bundle-/flower-like particles having different sizes were observed for synthesis using an in situ method. The UV protection ability of ZnO NPs coated on textiles was investigated using UV-Vis spectroscopy by measuring the UV protection factor (UPF) in the range of 280–400 nm. Higher values of UPF were obtained for ZnO NPs prepared using an in situ method. The UPF value obtained by this method was found to be 320, which demonstrates its excellent ability to block UV radiation. The antibacterial activities of ZnO NPs synthesized by the two methods possess very good bacteriostatic activity against Staphylococcus aureus and Escherichia coli bacteria demonstrated by the zone of inhibition.