Detection of human factor B biosynthesis in culture supernatants and cell lysates by ELISA.
We report here on the development of a simple, sensitive, convenient, and quantitative enzyme-linked immunosorbent assay for human factor B, a protein of the alternative complement pathway, by the sandwich method using goat anti-human factor B antibody. The assay described herein is reproducible and highly specific for human factor B. The assay was used to determine biosynthesis (cell lysates or extracts) and secretion (supernatants) of human factor B using human monocyte cell line U937. Phorbol myristate acetate strongly enhanced (10- to 20-fold) biosynthesis of factor B by U937. The combination of a sensitive enzyme-linked immunosorbent assay and phorbol myristate acetate enabled us to use a microculture system.
Mitani
M
M
All Children's Hospital, University of South Florida, St. Petersburg.
Good
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Day
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D000963:Antimetabolites / Q000494:pharmacology; D001345:Autoradiography; D002478:Cells, Cultured; D011415:Complement Factor B / Q000096:biosynthesis*; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D016685:Mitomycin; D008937:Mitomycins / Q000494:pharmacology; D009000:Monocytes / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000963:Antimetabolites; D008937:Mitomycins; D003609:Dactinomycin; D016685:Mitomycin; D003513:Cycloheximide; D011415:Complement Factor B; D013755:Tetradecanoylphorbol Acetate