How to easily detect luciferase activity in cell based assays?

Luciferase is the general term given to a class of oxidative enzymes which catalyze reactions that give off light, a process known as bioluminescence (Fig. 1).

structure and mechanism

Fig. 1: Mechanisms of the assay

In biology, researchers take advantage of this reaction to 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. Especially in screening processes it is crucial that the assay be accomplished in a single step, meaning the lysis buffer has to be formulated with D-luciferin.

One-Step Luciferase Assay System

BPS Biosciences has developed such a one-step luciferase assay system, which allows the reading of bioluminescence on a luminometer after a brief incubation period.

The One-Step Luciferase Assay System is designed to be used for high-throughput, sensitive quantitation of firefly luciferase activity in mammalian cell culture. The reagent consists of two components:

  • Luciferase Reagent Buffer (Component A)
  • Luciferase Reagent Substrate (Component B)

    Comparison with competitor

    Fig. 2: Comparison of the sensitivity of BPS’ One-Stop Luciferase Reagent with the main competitor’s reagent

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 benefits:

  • Sensitive – highly sensitive detection of firefly luciferase activity (Fig. 2)
  • Stable – the signal output is stable for more than two hours, providing flexibility with regard to incubation time  (see Fig. 3)
  • Convenient and HTS compatible – simple one-step, homogenous protocol
  • Compatibility – works well with a variety of common media containing 0-10% serum and phenol red
  • Instrumentation – does not require a luminometer with injectors

    Stability

    Fig.3: Stability of the One-Stop Luciferase Reagent

Interested in an price attractive alternative for the reagents you are used to? You might be interested in learning more, ask your questions and leave any comments below.

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