Versatile method to detect luciferase in reporter cell lines

luciferase-formula

Fig 1: How does Luciferase work?

Luciferase is the general term given to a class of oxidative enzymes that catalyze reactions that give off light, a process known as bio-luminescence (Fig. 1). In biology, researchers can take advantage of this reaction and use it as a readout for various biological processes. This has perhaps been exploited most in luciferase reporter cell lines where a promoter region from a gene of interest is placed immediately upstream of the coding sequence for luciferase. In this system, transcriptional activation of the gene of interest leads to a level of luciferase expression that is proportional to the level of gene activation.

The level of activation is then assessed by lysing the cells and adding the luciferase substrate, D-luciferin.

sensitivity

Fig 2: Sensitivity of the One-Step Luciferase Assay

BPS Bioscience has developed the One-Step Luciferase Assay System, which is accomplished in a single step as the lysis buffer has been formulated with D-luciferin. After a brief incubation period, bioluminescence is read on a luminometer. The assay is designed to be used for high-throughput, sensitive quantitation of firefly luciferase activity in mammalian cell culture. The reagent consists of two components, a Luciferase Reagent Buffer (Component A) and Luciferase Reagent Substrate (Component B). Component A and Component B are combined to form a working solution that contains all the necessary components for cell lysis and luciferase quantitation. This assay system has several features:

 

stability

Fig 3: The signal from the ONE-Step Luciferase reagent is strong and stable for more than 2 hours at RT.

  • Sensitivity – highly sensitive detection of firefly luciferase activity (see comparison with competitor’s reagent in Fig 2).
  • Stability – the signal output is stable for more than two hours, providing flexibility with regard to incubation time (Fig 3).
  • Convenience – simple one-step, homogeneous protocol.
  • High-throughput – one-step homogeneous protocol minimizes handling steps to support high-throughput screening applications.
  • Compatibility – works well with a variety of common media containing 0-10% serum and phenol red.
  • Instrumentation – does not require a luminometer with injectors.

 

nfkappa-b-cell-line-results

Fig 4: IL-1β dose response in NF-kappaB reporter (Luc)-HEK293 cells.

tgfs-cell-line-results

Fig 5: Dose response of SBEreporter-HEK293 cells to human TGFβ1.

The One-Step Luciferase Assay can be used with a wide variety of reporter cell lines provided by BPS Biosciences as well as other reporter cell lines based on firefly luciferase reporters.

Here is a selection of reporter cell lines available:

 

Are you interested in testing the One-Step Luciferase Assay in your laboratory? Contact me with the form below, I’ll be pleased to provide you with a trial quote.

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Written by Ali El Baya, PhD
Ali el Bayâ is the Sales Manager at tebu-bio for the North of Europe.