INDIGlo Luciferase Detection Reagent Kit
|Product Family||Product Number||Product Description||Technical Manual|
|LDR||LDR-10||10 mL each of 2x-concentrated INDIGlo LDR and Dilution Buffer||Technical Manual|
|LDR-25||25 mL each of 2x-concentrated INDIGlo LDR and Dilution Buffer||Technical Manual|
|LDR-50||50 mL each of 2x-concentrated INDIGlo LDR and Dilution Buffer||Technical Manual|
Bulk volumes of INDIGlo Luciferase Detection Reagent can be manufactured and packaged at any scale to meet customers’ specific requirements.
INDIGlo Luciferase Detection Reagent
INDIGlo Luciferase Detection Reagent (LDR) is formulated to deliver steady, long-lived luminescence signal from mammalian reporter cells expressing firefly luciferase. For convenience, the reagent is provided ‘ready to use’, without the requirement to first re-constitute powder-form components.
INDIGlo LDR works equally well in quantifying expressed luciferase activity from 96- or 384-well format reporter assays. The resulting light emission is stable over a 2 hr period (Figure 1), thereby making INDIGlo LDR an ideal, economical detection reagent for both low- and high-throughput batch processing of assay plates.
INDIGlo LDR is formulated as a 2x-concentrated reagent, thereby allowing greater versatility in its use. The 2x-concentrated reagent will be used when using 384-well or 1536-well assay plates that are processed using a homogenous assay strategy.
Alternatively, when processing 96-well assay plates either a homogenous or non-homogenous assay strategy may be used. For convenience, this kit provides an optimized Dilution Buffer for use in diluting INDIGlo LDR when performing non-homogenous assays.
Regardless of one’s assay setup preference INDIGlo LDR delivers rapid lysis of the reporter cells, robust luminescence signal, and sensitive quantification of expressed luciferase activities.
INDIGlo Luciferase Detection Reagent Performance
INDIGlo LDR performance from both Homogenous and Non-Homogenous assays. Dose-response analyses of Human AhR (INDIGO #IB06001, 96-well assay) was performed using the reference agonist MeBIO. AhR reporter cells were plated across two white assay plates, MeBIO treatment media were added, and the assay plates incubated for 24 hr. Plates were then processed with luciferase detection reagents in either a.) a homogenous or b.) a non-homogenous assay format. Resulting assay metrics are also shown from using another extended-glo detection reagent (Vendor X).
INDIGlo LDR potency is stable following repeated freeze and thaw cycles. INDIGlo LDR was frozen at -20°C and thawed either 1, 2 or 3 times, then used to quantify luciferase activity from HEK-293:luc cells cultured in a white 96-well assay plate. No loss of reagent potency is observed between these conditions.
INDIGlo Luciferase Detection Reagent Components and Storage Conditions
INDIGlo Luciferase Detection Reagent kits provide two reagents in equal volumes:
- INDIGlo LDR, formulated as a 2x-concentrated reagent
- Dilution Buffer, for optional use if performing non-homogenous assays
INDIGlo LDR is packaged in amber tubes, sealed under argon, then frozen. Upon receipt, both reagents should be further stored at -20°C. INDIGlo LDR is stable for at least 6 months with the expiration date printed on the Product Qualification Insert that is enclosed in each kit.
After INDIGlo LDR has been thawed for the first time, it may be subjected to an additional three freeze / thaw cycles without loss of potency. If more than three thaw-cycles are anticipated, it is advised to dispense the first-time thawed reagent into smaller single-use aliquots, which are then stored frozen.