Reconstitution of BioLux Cypridina Luciferase Substrate (E3314)

Introduction

IMPORTANT: 
Lyophilized substrate can be stored at -20°C for up to 1 year without loss of activity. 

Reconstituted substrate should be prepared only if you plan to perform the assays in the near future. 

Aliquots of reconstituted substrate can be stored at -80°C for up to 2 months.

Protocol

1. Preparation of Reconstituted Substrate (100X solution):
    Briefly centrifuge the thawed BioLux Cypridina Substrate Solvent vial at 12,000 rpm for 30 seconds.
2. Add 0.5 ml of absolute ethanol (not included) to the BioLux Cypridina Substrate Solvent and mix well.
3. Carefully lift the rubber cap of the lyophilized BioLux Cypridina Substrate vial and add 60 μl of the ethanol-solvent mixture (for small kit) or 600 μl of the ethanol-solvent mixture (for large kit) to the lyophilized BioLux Cypridina Luciferase Substrate.
4. Gently mix to dissolve the substrate (do not vortex, do not create bubbles by pipeting) and incubate at 4°C for 10–15 minutes (protect from light).
5. Gently mix again and incubate at 4°C for an additional 10–15 minutes (protect from light). 
   
   Note: Be sure to dissolve the residual lyophilized powder on the rubber cap.
6. Aliquot the reconstituted substrate (100X solution) and store at -80°C.