TECHNICAL CONSIDERATIONS IN THE PREPARATION OF FLUORESCENT-ANTIBODY CONJUGATES.

Applied microbiology ·Vol. 12 ·1964-07-00 ·Pages 343-8

LEWIS VJ, JONES WL, BROOKS JB, CHERRY WB

Abstract

A comparison was made of (NH(4))(2)SO(4), HCl, ethodin, and ethanol for fractionation of rabbit antiserum prior to conjugation with fluorescein isothiocyanate. Fractionation with the salt was found to be the method of choice from the standpoints of simplicity and recovery of antibody effective in conjugates prepared from the fractions. Effects of pH, temperature, dye-protein ratio, and molarity and type of buffer upon conjugation were studied. These technical factors were adjusted to produce conjugates for Corynebacterium diphtheriae which possessed higher specific titers than did reagents obtained by previously employed techniques.

Keywords

BLOOD PROTEIN ELECTROPHORESIS CORYNEBACTERIUM DIPHTHERIAE ESCHERICHIA COLI EXPERIMENTAL LAB STUDY FLUORESCEINS FLUORESCENT ANTIBODY TECHNIC IMMUNE SERUMS KLEBSIELLA RABBITS SPECTROPHOTOMETRY THIOCYANATES

MeSH Terms

Animals Blood Protein Electrophoresis Corynebacterium diphtheriae Escherichia coli Fluoresceins Fluorescent Antibody Technique Humans Immune Sera Indicators and Reagents Klebsiella Lagomorpha Proteins Rabbits Research Spectrophotometry Thiocyanates

Chemicals

Fluoresceins Immune Sera Indicators and Reagents Proteins Thiocyanates

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

LEWIS V J

JONES W L

BROOKS J B

CHERRY W B

References (10)

10 references, click to expand

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IDENTIFICATION OF CORYNEBACTERIUM DIPHTHERIAE WITH FLUORESCENT ANTIBACTERIAL REAGENTS.
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Superiority of fluorescein isothiocyanate (Riggs) for fluorescent-antibody technic with a modification of its application.
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Isothiocyanate compounds as fluorescent labeling agents for immune serum.
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A comparison of fluorescein-labeled gamma-globulins purified by Rivanol and DEAE chromatography.
J Immunol. 1963 Jan;90:116-20 PMID: 13945817
The staining properties of human serum proteins conjugated with purified fluorescein isothiocyanate.
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Fluorescent-Antibody Identification of Group A Streptococci from Throat Swabs.
Am J Public Health Nations Health. 1963 Jul;53(7):1083-92 PMID: 18017996
Fluorescent-antibody technic for serodiagnosis of Trypanosoma cruzi infection.
Proc Soc Exp Biol Med. 1959 Jul;101(3):540-3 PMID: 13675310
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Article Info

Journal

Applied microbiology

Abbr.

Appl Microbiol

ISSN

0003-6919

DOI

10.1128/am.12.4.343-348.1964

Published

1964-07-00

Pages

343-8

Language

English

Region

United States

NLM ID

7605802

PMCID

PMC1058130

Subset

External Links

PubMed source DOI: 10.1128/am.12.4.343-348.1964 PMC Full Text

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