Monoclonal antibodies against eucaryotic ribosomes. Use to characterize a ribosomal protein not previously identified and antigenically related to the acidic phosphoproteins P1/P2.
Mice were immunized against chick ribosomes with the use of various protocols and immunogen preparations. Hybridomas were prepared, clones screened, and specific antibodies identified by reversible protein staining followed by immunoperoxidase staining on nitrocellulose blots. Clones were obtained which secreted specific antibodies against ribosomal proteins S6, L7, L18a, P1/P2, and also against ribosomal RNA. Antibodies were typed by means of a dot-binding assay with typing antibodies immobilized on a solid support of nitrocellulose, and also characterized by their species cross-reactivities. The common determinant on proteins P1 and P2 cross-reacted with proteins of similar molecular weight in all eucaryotes tested, and with a determinant in a previously uncharacterized 38,000-dalton protein of the large ribosomal subunit. We designate this protein P0. The determinant of P0 was also present in a protein of similar molecular weight in all eucaryotes tested. Unlike P1 and P2, P0 was not removable from ribosomes by an ethanol-NH4Cl washing procedure. No evidence for a precursor-product relationship between P0 and P1/P2 was found. P0, P1, and P2 were found in active polysomes and in the nucleolus. The molecular weights of the nucleolar forms were not identical with those of the cytoplasmic forms, suggesting some processing during ribosomal assembly and/or transport.
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002459:Cell Fusion; D002645:Chickens; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010750:Phosphoproteins / Q000276:immunology*; D010954:Plasmacytoma; D012269:Ribosomal Proteins / Q000276:immunology*; D012270:Ribosomes / Q000276:immunology*; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000939:Epitopes; D010750:Phosphoproteins; D012269:Ribosomal Proteins