Single-step purification of epoxide hydrolase from rat liver microsomes using monoclonal-antibody chromatography.
Monoclonal antibody to rat liver microsomal epoxide hydrolase has been obtained after immunization of mice with the enzyme prepared by conventional methods. Antibody from ascitic fluid was purified and coupled to CNBr-activated Sepharose 4B, to give a specific affinity column. Monoclonal-antibody affinity chromatography provided a rapid single-step method of purifying to homogeneity active epoxide hydrolase from crude solubilized microsomes. The techniques used offer an effective method for characterization of a non-inhibitory monoclonal antibody.
D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity / Q000379:methods; D004851:Epoxide Hydrolases / Q000276:immunology / Q000302:isolation & purification*; D007120:Immunochemistry / Q000379:methods; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008862:Microsomes, Liver / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D004851:Epoxide Hydrolases