Development and testing of radio and enzyme immunoassays for acidic fibroblast growth factor (aFGF)
Autor: | Béatrice Groux-Muscatelli, José Courty, Jacques Grassi, Philippe Pradelles, Jean-Pierre Caruelle, Danièle Caruelle, Denis Barritault |
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Rok vydání: | 1988 |
Předmět: |
Immunoblotting
Basic fibroblast growth factor Radioimmunoassay Biophysics Enzyme-Linked Immunosorbent Assay Fibroblast growth factor Biochemistry Immunoenzyme Techniques Iodine Radioisotopes Mice chemistry.chemical_compound Antibody Specificity medicine Animals Molecular Biology Polyacrylamide gel electrophoresis Cells Cultured chemistry.chemical_classification Detection limit medicine.diagnostic_test biology Cell Biology Molecular biology Fibroblast Growth Factors Enzyme chemistry Polyclonal antibodies Immunoassay Antibody Formation Acetylcholinesterase biology.protein Cattle Electrophoresis Polyacrylamide Gel Female Immunization Rabbits Mitogens |
Zdroj: | Analytical Biochemistry. 173:328-339 |
ISSN: | 0003-2697 |
DOI: | 10.1016/0003-2697(88)90197-2 |
Popis: | Acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor from bovine brain stimulate growth in a variety of tissues in several species. Despite the 55% amino acid sequence homology of the two forms of FGF, a specific immunoassay of aFGF has been developed using a polyclonal antibody raised in a rabbit. Two immunoassays were compared: a radioimmunoassay (RIA) using 125I aFGF and an enzyme immunoassay (EIA) using aFGF coupled to the tetrameric form of acetylcholinesterase (aFGF-AchE) as tracer. With EIA, the detection limit was 1.5 ng/ml, versus 2.2 ng/ml with RIA, while the dose at 50% was 5.9 ng/ml for EIA and 9.6 ng/ml for RIA. Using a modified EIA procedure where aFGF-AchE was added 2 h after the other reagents, the dose at 50% binding was 1.5 ng/ml. Examples of the performance of both immunoassays are presented for various brain extracts of different species including human. The aFGF content obtained by these methods correlates (CR = 0.987) with the values obtained by biological assay. |
Databáze: | OpenAIRE |
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