A sensitive method of detecting proteins on dot and Western blots using a monoclonal antibody to FITC.
Monoclonal antibodies to FITC were produced and shown to be specific for the fluorochrome. Molecular weight marker proteins labelled with FITC could be detected after SDS-PAGE and transfer onto nitrocellulose using anti-FITC followed by an anti-mouse IgG-alkaline phosphatase conjugate. The molecular weight of an antigen common to Legionella pneumophila and recognised by a monoclonal antibody could be determined accurately on a Western blot when FITC labelled markers were used as internal standards. The FITC-anti-FITC system was shown to be extremely sensitive, detecting 23.7 amol of BSA-FITC conjugate (equivalent to 1.42 x 10(7) molecules of FITC) in a dot blot assay.
D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000942:Antigens, Bacterial / Q000032:analysis; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins / Q000276:immunology; D005456:Fluorescent Dyes; D007875:Legionella / Q000276:immunology; D008832:Microchemistry; D008970:Molecular Weight; D009113:Muramidase / Q000032:analysis; D010047:Ovalbumin / Q000032:analysis; D011485:Protein Binding; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000032:analysis; D013861:Thiocyanates / Q000276:immunology
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D005452:Fluoresceins; D005456:Fluorescent Dyes; D011506:Proteins; D013861:Thiocyanates; D012710:Serum Albumin, Bovine; D003101:Collodion; D010047:Ovalbumin; D009113:Muramidase; D016650:Fluorescein-5-isothiocyanate