Tech support: 877-222-9543

LightSwitch™ Luciferase Assay Kit
Catalog no. 32032 (100mL, 1000 assays)
OVERVIEWThe LightSwitch Luciferase Assay Reagent enables one-step reagent addition tomeasure luciferase reporter signal as part of the LightSwitch Luciferase Assay System.
The LightSwitch Luciferase Assay System is a fully optimized reporter system that
includes an improved luminescent reporter gene (RenSP), an optimized companion
assay reagent, and genome-wide collections of promoter and 3’UTR GoClone reporter
constructs (1). The GoClone reporter constructs have been optimized specifically foruse with the LightSwitch Luciferase Assay Reagent.

COMPONENTS & STORAGE
Qty

DescriptionStorage

100 mL

Assay Buffer-20°C for 6 months

1 tube
1 mL

PROTOCOL

Assay Substrate (100X) -20°C for 6 months
Substrate Solvent

room temp or -20°C

Note: Luciferase assays may be conducted immediately or the plates may be frozen
at -80˚C (freezing generally increases cell lysis and luciferase signal). If using frozen
plates, thaw and bring to room temperature before assaying.
1) Reconstitute 100X Substrate.
Add 1mL Substrate Solvent to tube of lyophilized Assay Substrate. Dissolve completely.
Protect from light and minimize time at room temperature. 100x Substrate may be
stored at -20C and protected from light for 2-3 weeks. For best results, use freshly
reconstituted substrate.

2) Prepare Assay Solution (for one 96-well plate).
Thaw 100mL bottle of Assay Buffer in room temperature water bath and add 1mLof reconstituted 100x Substrate just prior to use. Prepare Assay Solution (buffer +substrate mix) fresh for each use and use within 2-3 hours. To assay fewer wells,
make up only what you need and store remaining substrate and buffer separately at-20C. For best results, avoid additional freeze-thaw cycles. To thaw re-frozen buffer,incubate in a warm (37C) water bath for at least 1 hour and mix well to ensure that
all components go back into solution.
3) Add Assay Solution to wells.
Use a multi-channel pipettor to add 100uL Assay Solution (buffer + substrate)
directly to each sample well (100uL cells + media) in a white 96-well plate. The
Assay Buffer also serves as a lysis buffer. If cells were grown in another plate orflask, transfer samples to a white 96-well plate in 100uL total volume (media or PBS).

4) Incubate for 30 minutes at room temperature.
Cover plate and protect from light. If assaying more than one plate, stagger additionof Assay Solution so that each plate incubates for 30 minutes before reading.

5) Read each well for 2 seconds in a plate luminometer.
A recommended luminometer is the SpectraMax L (Molecular Devices)

Genome In. Function Out.

Tech support: 877-222-9543
This lot has passed our certificate of analysis requirements. More details upon request.
For suggested protocols and a complete description of the LightSwitch Luciferase
Assay System, see our resources page at http://switchgeargenomics.com/resources/(1) For research use only. This product and/or its use is subject to one or more of the following SwitchGear Genomics patents: U.S. Pat. Pub. Nos. US2007/0161031, US2009/0018031, US2008/0220983,
US61/397340 and various corresponding patent applications and issued patents. SwitchGear sells
licensed Renilla-based luciferase vectors. This product does not convey a license to use recombinant
Renilla luciferase under U.S. Pat. Nos. 5,292,658, 5,418,155 and related patents. Certain applications of
this product may require licenses from others.

Genome In. Function Out.