Development of a monoclonal antibody to the rabbit 8.5S uterine progestin receptor.

摘要: Nonactivated (8.5S) rabbit uterine progestin receptor was enriched 10- to 30-fold by chromatography on columns of spheroidal hydroxylapatite and DEAE-cellulose. A total approximately 25 micrograms (purity 1%) injected at multiple sites into a BALB/c mouse. After several injections, splenic lymphocytes were fused with P3x63Ag8.653 mouse myeloma cells. This fusion produced in excess 240 hybridomas, which screened an enzyme-linked immunosorbent assay (ELISA), solid-phase radioimmunoassay, sucrose gradient centrifugation. One colony (KN 382/EC1) immunoglobulin G1 bound 8.5S receptor. The cell line has been repeatedly cloned under conditions limiting dilution expanded mice as ascitic tumors. Antibody purified (NH4)2SO4 precipitation, DEAE-cellulose chromatography, affinity protein - Sepharose CL-4B. Specificity the antibody determined centrifugation radioimmunoassay. receptors from uterus MCF-7 breast cancer It did not react rat uterus, guinea pig or chick oviduct, nor it bind estrogen any tissues we tested.