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PMID: 96441 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Immunological screening method to detect specific translation products.

Broome S, Gilbert W

Abstract

We describe a very sensitive method to detect as antigens the presence of specific proteins within phage plaques or bacterial colonies. We coat plastic sheets with antibody molecules, expose the sheet to lysed bacteria so that a released antigen can bind, and then label the immobilized antigen with radioiodinated antibodies. Thus, the antigen is sandwiched between the antibodies attached to the plastic sheet and those carrying the radioactive label. Autoradiography then shows the positions of antigen-containing colonies or phage plaques. A few molecules of antigen released from each bacterial cell generatean adequate signal.

MeSH Terms
Antibody Specificity Bacteriolysis Dose-Response Relationship, Immunologic Escherichia coli Galactosidases/immunology Immunologic Techniques Lactose Factors Lysogeny Protein Biosynthesis Radioimmunoassay/methods beta-Galactosidase/biosynthesis,genetics,immunology
Chemicals
Galactosidases beta-Galactosidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Broome S
Gilbert W
References (5)
5 references, click to expand
  1. Immunoaffinity chromatography of proteins.
    Methods Enzymol. 1974;34:723-31 PMID: 4141422
  2. High-level production of -galactosidase by Escherichia coli merodiploids.
    J Bacteriol. 1972 Nov;112(2):856-60 PMID: 4563980
  3. Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film.
    FEBS Lett. 1977 Oct 15;82(2):314-6 PMID: 913604
  4. In situ immunoassays for gene translation products in phage plaques and bacterial colonies.
    Gene. 1976;1(1):65-79 PMID: 829886
  5. Solid-phase radioimmunoassay in antibody-coated tubes.
    Science. 1967 Dec 22;158(3808):1570-2 PMID: 6060361
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1978-06-00
Pages
2746-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC392640
Subset
IM
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