The objective of this work was to study the effect of temperatures and incubation times on Silk Fibroin (SF)/Chitosan (CS) blend films properties. The films were firstly prepared by mixing the SF and CS solution in homogeneously blended, then cast on the polystyrene culture plates before taking into the oven at 40°C for 3 days. The obtained SF/CS blend films were treated at 90 and 120°C, each for 30, 60 and 90 min, respectively. All of films were characterized for their morphology and secondary structures by using SEM and FTIR, respectively. The results showed that SF/CS films have homogeneous texture without phase separation. However, they also appeared some particles dispersed and embedded on their surfaces. The surfaces of the films slightly increased smooth texture when increased both temperatures and incubation times. The results were also noted that the characteristics were varied by those different temperatures. FTIR spectra indicated that the absorption bands at amide regions (I, II and III) of the SF were similar profile. Moreover, treatment the blended films with temperatures resulted to changed the secondary structures of the films. This means the ratio of β-sheet were increased. It can be expected that water evaporation by increasing temperature resulted to make the molecules of SF and CS close up and help to form H-bonds between them.