Preparation and properties of a cytotoxic monoclonal rat anti-mouse Thy-1 antibody.
A hybrid cell line secreting a cytotoxic rat IgM antibody with specificity for both the 1 and 2 alleles of mouse Thy-1 was selected. The fusion was between rat spleen cells immunised to SBA spleen cell plasma membranes and the rat myeloma cell line 210RCY3-Ag123. The cell line can be propagated in Lou-strain rats to yield large volumes of antibody-containing ascites with a cytotoxic titre of 10(6)-10(7) for peripheral T cells. The antibody is relatively stable and can be used for indirect fluorescence assays in conjunction with a fluorochrome (e.g. fluorescein)-coupled goat anti-rat immunoglobulin (pre-absorbed on mouse immunoglobulin). More simply, it can be directly conjugated to a fluorochrome and used in association with an appropriately absorbed goat anti-mouse Ig coupled with a contrasting and different fluorochrome (e.g. rhodamine) to allow simultaneous enumeration of both T and B lymphocytes.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000954:Antigens, Surface / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D005455:Fluorescent Antibody Technique; D006041:Goats; D006168:Guinea Pigs; D007075:Immunoglobulin M / Q000096:biosynthesis; D008565:Membrane Proteins / Q000276:immunology*; D051379:Mice; D008806:Mice, Inbred AKR; D008808:Mice, Inbred CBA; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D013601:T-Lymphocytes / Q000276:immunology; D018800:Thy-1 Antigens
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D007075:Immunoglobulin M; D008565:Membrane Proteins; D018800:Thy-1 Antigens