Abstract: Modified sol–gel processed silica matrices were prepared to explore the possibilities of using these matrices for electrophoresis applications. The sol–gel was prepared at room temperature using tetraethyl orthosilicate as a precursor at different molar ratios (R = 4, 16, 32 and 64) in the presence of sodium phosphate buffer. The sol–gel derived from R = 32 formed smooth and transparent gel, and showed high transmittance value (97%) compared to other molar ratios. Transmission electron microscopy and scanning electron microscopy studies of gel matrix (R-32) confirmed randomly distributed interstitial space of variable size (30–60 nm) with slit like openings on the gel surface. The R = 32-based matrix internal and surface characteristics were suitable for gel electrophoresis. Therefore, sol–gel electrophoresis was carried out using gel matrix (R = 32) on molecular markers and pre-stained proteins (08–220 kDa). The silica gel matrix allowed migration as well as separation of proteins based on their molecular weights. The results were compared with standard gels (agarose and polyacrylamde). The aim of the work to prepare a silica matrix suitable for gel electrophoresis which could be utilised to develop electrophoresis-based point-of-care technologies for clinical diagnostics. Graphical Abstract: Schematic illustration of sol–gel electrophoresis (SoGE), showing different stages of sol–gel preparation and its utilisation for separation of proteins 8–220 kDa. © 2017, Springer Science+Business Media New York.