Abstract
A rapid, reproducible, and relatively simple method for the preparation of large quantities of fluorescein isothiocyanate-conjugated antibody is presented. Tagged globulins with fluorochrome-protein ratios of 1 and 2 were eluted from diethylaminoethyl-cellulose in batch form by using sodium phosphate-buffered 0.1 M and 0.2 M saline, respectively. The intensity and specificity of the fluorescence due to this antibody was quite high, as determined by the ability of untagged antibody to block the immunological reactivity of the fluorescent antibody and by the lack of fluorescence of antigenically unrelated material when treated under identical conditions.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- DEDMON R. E., HOLMES A. W., DEINHARDT F. PREPARATION OF FLUORESCEIN ISOTHIOCYANATE-LABELED GAMMA-GLOBULIN BY DIALYSIS, GEL FILTRATION, AND IONEXCHANGE CHROMATOGRAPHY IN COMBINATION. J Bacteriol. 1965 Mar;89:734–739. doi: 10.1128/jb.89.3.734-739.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- KAUFMAN L., KAPLAN W. Preparation of a fluorescent antibody specific for the yeast phase of Histoplasma capsulatum. J Bacteriol. 1961 Nov;82:729–735. doi: 10.1128/jb.82.5.729-735.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
- KAUFMAN L., KAPLAN W. SEROLOGICAL CHARACTERIZATION OF PATHOGENIC FUNGI BY MEANS OF FLUORESCENT ANTIBODIES. I. ANTIGENIC RELATIONSHIPS BETWEEN YEAST AND MYCELIAL FORMS OF HISTOPLASMA CAPSULATUM AND BLASTOMYCES DERMATITIDIS. J Bacteriol. 1963 May;85:986–991. doi: 10.1128/jb.85.5.986-991.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lewis V. J., Brooks J. B., Jones W. L. Comparison of concentration methods for fluorescent antibody reagents. Am J Med Technol. 1966 May-Jun;32(3):201–205. [PubMed] [Google Scholar]
- Olson W. P. Maximal brightness and yield of fluorescent antibody. J Bacteriol. 1968 Mar;95(3):1176–1177. doi: 10.1128/jb.95.3.1176-1177.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RIGGS J. L., SEIWALD R. J., BURCKHALTER J. H., DOWNS C. M., METCALF T. G. Isothiocyanate compounds as fluorescent labeling agents for immune serum. Am J Pathol. 1958 Nov-Dec;34(6):1081–1097. [PMC free article] [PubMed] [Google Scholar]
- Wood B. T., Thompson S. H., Goldstein G. Fluorescent antibody staining. 3. Preparation of fluorescein-isothiocyanate-labeled antibodies. J Immunol. 1965 Aug;95(2):225–229. [PubMed] [Google Scholar]