This article reports the effects of gamma irradiation (dose ranges 0.1–10 kGy from 60Co source) on the characteristics of solid gelatin and the physico-mechanical, microstructural and bioactive properties of the scaffold prepared from irradiated gelatin solution. FTIR, intrinsic viscosity, bloom strength, thermal properties, SEM, tensile properties, water uptake ability and antimicrobial activities of non-irradiated and irradiated solid gelatin and its scaffolds were investigated. The detailed experimental results for the solid gelatin demonstrated that 1 kGy γ-irradiated samples showed higher intrinsic viscosity, enhanced thermal stability and bloom strength than other irradiated samples. Furthermore, the scaffold thus prepared from irradiated and non-irradiated gelatin also revealed that 1 kGy samples showed the highest tensile strength and modulus with good water resistivity than other irradiated and non-irradiated samples. In addition to the physico-mechanical properties, 1 kGy scaffolds have also exhibited the highest resistivity towards microbial growth that can have potentiality as scaffold in biomedical sector. The enhanced functional and bioactive properties at low irradiation doses (1 kGy) may occurred due to an initial breaking of hydrogen bonds of polypeptide chains in gelatin molecules that indicated by the shift of amide A, I and II peaks to higher wave numbers in FTIR. This enhancement resulted probably due to the domination of crosslinking over degradation at 1 kGy. It was also observed that 1 kGy γ-radiation-induced crosslinking has lowered the hydrophilicity by decreasing water uptake and mean pore diameter of the interconnected porous structures of gelatin.