High-Throughput Firefly Luciferase Reporter Assays

Methods Mol Biol. 2018:1755:19-29. doi: 10.1007/978-1-4939-7724-6_2.

Abstract

Firefly luciferase reporter gene assays find wide application in high-throughput screens to identify molecular components of biological networks or to identify chemical compounds capable of interfering with cellular signaling. Here, we present methods to prepare affordable firefly luciferase assay reagents and procedures to use these reagents in reporter gene high-throughput screening with large batches of 96-well cell culture plates.

Keywords: Assay reagent; Firefly luciferase; High-throughput screening; Reporter gene assay; Stable luminescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Assay / instrumentation
  • Biological Assay / methods*
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Cell Line
  • Firefly Luciferin / chemistry
  • Genes, Reporter / genetics*
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods*
  • Humans
  • Luciferases, Firefly / chemistry
  • Luciferases, Firefly / genetics*
  • Luciferases, Renilla / chemistry
  • Luciferases, Renilla / genetics
  • Luminescent Agents / chemistry
  • Luminescent Measurements / instrumentation
  • Luminescent Measurements / methods
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism

Substances

  • Luminescent Agents
  • RNA, Small Interfering
  • Firefly Luciferin
  • Luciferases, Renilla
  • Luciferases, Firefly