A simplified microassay for inhibition of protein synthesis in reticulocyte lysates by immunotoxins

Oliver W. Press, Ellen S. Vitetta, Paul J. Martin

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We describe a simplified cell free reticulocyte lysate protein synthesis assay which has been shown to reliably demonstrate dose dependent inhibition of protein synthesis with cycloheximide and with four ricin A chain immunotoxin conjugates. Maximal [3H]leucine incorporation was observed using 40 μl of reticulocyte lysate per microtiter well and by incubating samples for 30 minutes before harvesting with a multiple automated sample harvester. Trichloroacetic acid precipitation and hydrogen peroxide decolorization were not required with this method. This rapid, simplified assay system will facilitate testing of agents that inhibit protein synthesis, particularly immunotoxins.

Original languageEnglish (US)
Pages (from-to)37-41
Number of pages5
JournalImmunology Letters
Volume14
Issue number1
DOIs
StatePublished - Nov 17 1986

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Immunotoxins
Reticulocytes
Ricin
Trichloroacetic Acid
Proteins
Cycloheximide
Leucine
Hydrogen Peroxide

Keywords

  • Immunotoxin
  • Protein synthesis
  • Reticulocyte Lysate
  • Ricin
  • Toxin

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

A simplified microassay for inhibition of protein synthesis in reticulocyte lysates by immunotoxins. / Press, Oliver W.; Vitetta, Ellen S.; Martin, Paul J.

In: Immunology Letters, Vol. 14, No. 1, 17.11.1986, p. 37-41.

Research output: Contribution to journalArticle

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