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
- Preparation of Reconstituted Substrate (100X solution):
Briefly centrifuge the thawed BioLux Cypridina Substrate Solvent vial at 12,000 rpm for 30 seconds. - Add 0.5 ml of absolute ethanol (not included) to the BioLux Cypridina Substrate Solvent and mix well.
- 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.
- 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).
- 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. - Aliquot the reconstituted substrate (100X solution) and store at -80°C.