2022 Volume 66 Issue 1 Pages 5-11
A complex of carboxylesterase and transferrin was obtained after cytosolic proteins in the mouse liver were separated by a combination of non-denaturing two-dimensional electrophoresis (2DE) and reversible staining. At the site where the complex was separated on the 2DE gel, the complex retained esterase activity because the relative intensity of the fluorescent esterase substrate was 1.6-fold higher than that within the 2DE gel in the absence of protein. Furthermore, the complex was extracted from the 2DE gel and bound to antibody-conjugated protein G plus A (protein A/G) agarose by non-denaturing electrophoresis after separation and detection. The esterase activity was retained after the complex was extracted and bound to antibody-conjugated protein A/G agarose. These results indicate that intact protein complexes were separated, detected, and extracted by non-denaturing electrophoresis.