ANTIBODIES TO REDUCED GLUTATHIONE

Reduced glutathione was conjugated to carrier proteins with glutaraldehyde. Conjugates were reduced by sodium borohydride and injected into rabbits. Using an enzyme-linked immunosorbent assay, antibody affinity and specificity were determined by competition experiments between glutathione conjugate and related conjugated compounds. The resulting cross-reactivity ratios, calculated at half-displacement, showed that conjugated glutathione was the best recognized compound. Non-reduced glutathione conjugate was 50 x less recognized. The other related conjugates were not recognized at all. Thus, the high affinity and relative specificity make these antibodies potentially valuable tools for immunohistochemical detection of reduced glutathione in glutaraldehyde-fixed rat brain. Using purified antisera diluted at 1/5000, reduced glutathione was preferentially visualized in nerve fibers of cortex, cerebellum and spinal cord. These results suggest that concentration of GSH in rat CNS are higher in nerve fibers than in neuronal perikaryons.