ANTIBODIES TO REDUCED GLUTATHIONE

Reduced glutathione was conjugated to carrier proteins with glutaralde hyde. 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 gl utathione conjugate and related conjugated compounds. The resulting cr oss-reactivity ratios, calculated at half-displacement, showed that co njugated glutathione was the best recognized compound. Non-reduced glu tathione conjugate was 50 x less recognized. The other related conjuga tes were not recognized at all. Thus, the high affinity and relative s pecificity make these antibodies potentially valuable tools for immuno histochemical detection of reduced glutathione in glutaraldehyde-fixed rat brain. Using purified antisera diluted at 1/5000, reduced glutath ione was preferentially visualized in nerve fibers of cortex, cerebell um and spinal cord. These results suggest that concentration of GSH in rat CNS are higher in nerve fibers than in neuronal perikaryons.