INDIGO’s All-Inclusive Reporter Assays
Luciferase Reporter Assays
Our reporter assays capitalize on the extremely low background, high sensitivity, and broad linear dynamic range of bio-luminescence reporter gene technology. INDIGO’s reporter cells incorporate the cDNA encoding beetle luciferase, a 62 kD protein originating from the North American firefly (Photinus pyralis). Luciferase catalyzes the mono-oxidation of D-luciferin in a Mg+2-dependent reaction that consumes O2 and ATP as co-substrates, and yields as products oxyluciferin, AMP, PPi, CO2, and photon emission. Luminescence intensity of the reaction is quantified using a luminometer and is reported in terms of Relative Light Units (RLU’s). Thus, quantifying changes in luciferase expression in the test sample-treated reporter cells provides a sensitive surrogate measure of changes in activity.
INDIGO’s Reporter Cells
INDIGO Biosciences’ assay feature engineered receptor-specific reporter cells prepared using our unique CryoMite™ process. This proprietary cryopreservation process, enables long-term preservation of our unique reporter cells, so we can ship our live-cell assays to you via overnight delivery, for your immediate use. Or, you can store the assay kits at -80°C. Once thawed, reporter cells typically present greater than 95% cell viability and are ready for immediate use. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
INDIGO’s Glow Luciferase Reporter Assay Kits
INDIGO’s luciferase reporter assay systems are engineered to provide optimal assay sensitivity and dynamic range when quantifying receptor activity. Following ligand-activation, the receptor complex acts to induce expression of the luciferase reporter. Upon addition of detection reagent, the intensity of light emission from the luciferase reaction can be directly correlated to the activation status of the specific receptor. All INDIGO reporter assay kits incorporate a detection reagent specially formulated to provide stable light emission between 5 and 90+ minute after initiating the luciferase reaction, thus allowing users the ability to dispense detection reagent into all assay wells prior to commencing activity measurements. This eliminates the need for a luminometer equipped with injectors and allows plates to be processed in batch, which dramatically reduces the start-to-finish read time of assay plates. Test compounds may be screened for agonist or antagonist activities against receptors and transcription factors expressed within the cytoplasm and nuclear environments of healthy, dividing mammalian cells.
ALL- INCLUSIVE REPORTER ASSAY KIT FORMATS
INDIGO Biosciences’ luciferase reporter assays are optimally configured for processing in 96- and 384-well format assay plates. Single plate kits (1x 96- and 1x 384-well) are offered as all-inclusive assay systems. Each kit includes luciferase reporter cells, an assay plate, all required reagents (optimized culture media, positive-control agonist, and detection reagent), as well as a detailed Technical Manual.
Bulk Reagent Packs are offered to accommodate the needs of HTS users. These products are scaled for 10x 96- and 10x 384-well plate assays and provide luciferase reporter cells, optimal culture media, positive-control ligand, luciferase detection reagent, and a detailed Technical Manual and Protocol Quick Guide. Bulk Reagent Packs do NOT include assay plates because HTS facilities typically stock large quantities of such plates.
Please note that, regardless of assay format, Luciferase Reporter Cells are single-use reagents. Once thawed, Reporter Cells may not be refrozen or reused. Assay kits provide sufficient extra volumes of all reagents to accommodate the needs of performing mechanical transfers and test compound dilutions. However, extra volumes of reporter cells and Luciferase Detection Reagent should be discarded after each assay set-up.
Five basic configurations of our luciferase reporter assay kits are offered:
- 3x 32 assays in a 96-well format plate.
This all-inclusive kit contains reporter cells and sufficient materials to perform three distinct groups of 32 assays. Each group will comprise 32 nuclear receptor assays, and will utilize four 8-well strips affixed to a 96-well frame (provided). If desired, however, reagents may be combined to perform either 64 or all 96 assay reactions at one time.
- 1x 96-well assay.
This all-inclusive kit format contains reporter cells and all additional materials needed and is intended for users wishing to dedicate one entire 96-well plate to performing nuclear receptor assays.
- 1x 384-well assay.
This all-inclusive kit format is intended for users wishing to dedicate one entire 384-well plate to performing nuclear receptor assays.
- 960 assay Bulk Reagent Pack for (10x) 96-well plates.
This kit format is intended to accommodate HTS applications in which a minimum of ten (10) 96-well plates will be dedicated to a single screening run. Assay plates are NOT supplied with this product. Hence, users must provide white, sterile, cell-culture treated 96-well assay plates.
- 3,840 assay Bulk Reagent Pack for (10x) 384-well plates.
This kit format is intended to accommodate HTS applications in which a minimum of ten (10) 384-well plates will be dedicated to a single screening run. Assay plates are NOT supplied with this product. Hence, users must provide white, sterile, cell-culture treated 384-well assay plates.
All Bulk Reagent Packs are manufactured on-demand, and require a maximal lead time of 12 business days for delivery. We welcome inquiries pertaining to the custom manufacture of alternative volume Bulk Reagent Packs. Please let us know how we may better help you achieve your research goals!
All Luciferase Reporter Assay System products are shipped on dry ice for overnight delivery anywhere within the United States.
HOW TO USE OUR KITS: A QUICK GUIDE VIDEO
Perform nuclear receptor assays with easy-to-use kits from INDIGO Biosciences. This short video provides a brief overview of what to expect when working with our kits, including unpacking and storing kit contents, setting up and performing the assays, and reviewing data and results.
*Please note, this video is for INDIGO’s non-pre-incubation assays. INDIGO’s pre-incubation assays involve minor but crucial additional Day 1 steps and different dispensed volumes. Always refer to your Technical Manual (included in all kit shipments) for the most up-to-date information and proper technique. Find out which kits require pre-incubatio