The glulam-cross laminated timber (CLT) composite floor is a type of prefabricated composite floor that integrates glulam beams and CLT slab into a unified structure using shear connectors. To investigate the bending performance of the glulam-CLT composite floor, the bending test was conducted on a full-scale composite floor under static load. The study comprehensively analyzed the failure mechanism, load–deflection behavior, interface slip and strain distribution of the glulam-CLT composite floor. The test results of the composite floor indicated that the failure mode was tensile fracture of the wood beam at the bottom. As the load increased, the deflection deformation of the mid-span beam exceeded that of the edge beam. When the load reached its ultimate limit, the deflection deformation of the mid-span beam increased by 14.4% compared to the edge beam. In the early loading phase, the strain distribution of the composite section satisfied the assumption of a plane section. However, the strain distribution deviated from this assumption with the increased load due to the relative slips between the glulam beam and CLT flange. To calculate the bending performance of the composite floor, the M-shaped section of the glulam-CLT composite floor was simplified as T-section composite beams. The linear-elastic method for determining the flexural rigidity and ultimate bearing capacity of the glulam-CLT composite floors was proved to be accurate and reliable. The findings provided valuable insights into the bending behavior of the CLT flange under load and emphasized the non-uniform stress distribution caused by shear lag effects.