Human placental steroid sulphatase--purification and monospecific antibody production in rabbits.
Human steroid sulphatase was purified 43-fold from placental microsomes using a four step procedure: solubilization with Miranol H2M, Bio-Gel A 1.5 m chromatography, column chromatofocusing and Sephadex G-75 chromatography. The purified enzyme that appeared electrophoretically homogeneous was used to immunize rabbits. Protein blotting demonstrated that the resulting antiserum mainly reacted with a polypeptide of 63 000 dalton, which is about the size of placental steroid sulphatase. The antiserum was freed from minor impurities by absorbing it to Sepharose 4B with immobilized antigens prepared from a steroid sulphatase deficient placenta.
D000818:Animals; D000906:Antibodies; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D011817:Rabbits / Q000276:immunology; D043266:Steryl-Sulfatase; D013429:Sulfatases / Q000302:isolation & purification* / Q000378:metabolism
D000906:Antibodies; D013429:Sulfatases; D043266:Steryl-Sulfatase