Establishment of dye-uptake method (A375 assay) for quantitative measurement of IL-1: correlation with LAF assay

Dev Biol Stand. 1988:69:93-101.

Abstract

The dye-uptake method (A375 assay) for quantitative measurement of biological activity or recombinant human interleukin-1 (IL-1) was established based on the cytostatic activity of IL-1 on human melanoma A375 cells. Although various factors, e.g., incubation temperature, inoculum cell size and cell growth phase, were found to influence the titration to some degree, the assay-to-assay fluctuation in the observed titers determined by this method could be made minimum after normalization with the observed titer of a standard IL-1. A good correlation was observed between A375 and LAF assays. Thus, A375 assay was shown to be a stable and reliable method and could be replaced with LAF assay for quantitative measurement of recombinant human IL-1.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Cell Division / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Humans
  • Interleukin-1 / analysis*
  • Interleukin-1 / pharmacology
  • Lymphocyte Activation*
  • Melanoma
  • Methods
  • Methylene Blue
  • Mice
  • Mice, Inbred C3H
  • Recombinant Proteins / analysis*
  • Recombinant Proteins / pharmacology
  • Spectrophotometry / methods
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology

Substances

  • Interleukin-1
  • Recombinant Proteins
  • Methylene Blue