Related Experiment Video
Updated: Aug 19, 2026
Isolation and Activation of Murine Lymphocytes
Published on: October 30, 2016
Purification and partial characterization of 200 kDa oncofetal antigen from radiation induced murine lymphocytic
1Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile 36688.
Abstract:
1. A 200 kDa glycoprotein (gp200) oncofetal antigen was purified from solubilized membranes of a radiation-induced murine lymphocytic lymphoma cell line (XR11-5T), grown in syngeneic RFM mice, by successive gel chromatography of the active fraction on lentil lectin agarose, Q- and S-Sepharose and Superose-12 using an FPLC system. 2. A murine monoclonal antibody 115, produced by the syngeneic immunization of adult male C57BL/6N mice with 12-day mouse fetal cells, was used in a slot blot antibody assay to follow up the active fractions. 3. The purified glycoprotein has a pI of 5.4. 4. Treatment of radiolabeled gp200 with neuraminidase caused a slight reduction in size due to the removal of sialic acid groups and a shift in pI to 6.3. 5. Treatment of gp200 with different glycosidases shows that gp200 is susceptible to N- and O-glycanase but not to endoglycosaminidase H. 6. On extraction of gp200 with Triton X-114 it partitions exclusively into the detergent-rich fraction consistent with being an integral membrane protein.
