To elucidate the effect of preheating temperature and calcium ions on the properties of cold-set soybean protein gel, Ca2+ induced gelation of soybean protein were investigated by rheological approaches, electrophoresis analysis, confocal scanning laser microscopy (CSLM) and surface hydrophobicity (S0). The results showed that, both CaCl2 concentration (20–40 mM) and preheating temperature (80–120 °C) took significant influence on the dynamic viscoelasticity of the gel samples. The bands distribution of samples preheated at 120 °C were different from the bands distribution of samples preheated below 100 °C in denature and native electrophoresis. The CSLM analysis showed that the gel became coarser as the Ca2+ increased. On the other hand, two different kinds of gel were shown under the same Ca2+ concentration: a bead-like structure (below 100 °C) and a filamentous structure (above 100 °C). According to fractal theory, weak-linked gel (α was >0.8) was formed when preheating below 100 °C, while a transition gel (α = 0.52–0.62) was obtained when preheating over 100 °C. All these results suggested that the preheating temperature influence the type of cold-set gel.