The use of tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes

Byron Batteiger, Wilbert J. Newhall V, Robert B. Jones

Research output: Contribution to journalArticle

349 Citations (Scopus)

Abstract

The determination of the immunoreactivity of protein antigens in complex mixtures has been greatly facilitated by combining their separation via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with electrophoretic transfer to nitrocellulose membrane (NCM), and probing of bound proteins with specific antisera. Methods using various buffers and blocking agents have been published, but no studies have been published which compare these methods with each other or with others of potential merit. We have performed such a comparative study using protein antigens from Chlamydia trachomatis and Neisseria gonorrhoeae. In addition, we describe a method that blocks unoccupied protein binding sites on NCM with the nonionic detergent Tween 20, rather than proteins. This system proved to be equivalent or superior to other methods evaluated in the detection of immunoreactive proteins, and permitted staining of the NCM for protein after immunological probing. Such staining allowed precise identification of immunoreactive proteins. In addition, individual stained proteins could be excised and assessed for bound antibody in an indirect radioimmunoassay.

Original languageEnglish
Pages (from-to)297-307
Number of pages11
JournalJournal of Immunological Methods
Volume55
Issue number3
DOIs
StatePublished - Dec 30 1982

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Collodion
Polysorbates
Membranes
Proteins
Staining and Labeling
Antigens
Neisseria gonorrhoeae
Chlamydia trachomatis
Complex Mixtures
Protein Binding
Sodium Dodecyl Sulfate
Detergents
Radioimmunoassay
Immune Sera
Polyacrylamide Gel Electrophoresis
Buffers
Membrane Proteins
Binding Sites
Antibodies

Keywords

  • antigenic analysis
  • chlamydial proteins
  • immunoblot
  • indirect radioimmunoassay

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

The use of tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. / Batteiger, Byron; Newhall V, Wilbert J.; Jones, Robert B.

In: Journal of Immunological Methods, Vol. 55, No. 3, 30.12.1982, p. 297-307.

Research output: Contribution to journalArticle

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