Preparation of Luciferin for In Vitro Bioluminescent Assays


  • D-Luciferin Firefly, potassium salt, 1.0 g /vial

(Caliper Life Sciences Part Number #119222)

  • Sterile water
  • Complete media


  1. Prepare a 200X Luciferin stock solution (30 mg/ml) in sterile water. Mix gently by inversion until Luciferin is completely dissolved. Use immediately, or aliquot and freeze at -20 °C for future use.

Note: One can either reconstitute the entire 1.0 g of D- Luciferin in 33.3 mL of sterile water to make the 30 mg/mL (200x) stock solution, or reconstitute the quantity of D-Luciferin necessary for an individual experiment.

  1. Prepare a 150 μg/mL working solution of D-Luciferin in pre-warmed tissue culture medium.

Quick thaw 200X stock solution of Luciferin and dilute 1:200 in complete media (150 μg/mL final).

  1. Aspirate media from cultured cells.
  2. Add 1x Luciferin solution to cells just prior to imaging.

Note: Incubating the cells for a short time at 37 °C before imaging can increase the signal.

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