Aryl Hydrocarbon Receptor (AhR)
| Product Family | Product Number | Product Description | Technical Manual |
| IB0600 AhR |
IB06001-32 | Human AhR Reporter Assay System, 3 x 32 assays in 96-well format | Technical Manual |
| IB06001 | Human AhR Reporter Assay System, 1 x 96-well format assays | Technical Manual | |
| IB06002 | Human AhR Reporter Assay System, 1 x 384-well format assays |
Aryl Hydrocarbon Receptor Assay Kit
This Human Aryl Hydrocarbon Receptor (AhR) assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to AhR Reporter Cells, two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist (MeBIO), Luciferase Detection Reagent, and a cell culture-ready assay plate.
INDIGO’s cell-based reporter assays allow scientists to detect any biological activity that their test samples may exert against a specific receptor present in the cell. They utilize firefly luciferase reporter gene technology which provides superior precision and sensitivity. Since the receptor binding controls the expression of the luciferase reporter gene, luciferase activity in the cells can be correlated directly with the activity of the receptor. The strength of an interaction of a chemical with the target receptor is quantified using a luminometer to measure the level of light emitted.
Fast, reproducible, easy-to-analyze results are only four steps away
Many luciferase reporter assays require the user to grow their own cells and take time to optimize the results. INDIGO’s reporter cells contain the receptor of interest and the luciferase reporter gene. Reporter cells have been optimized to provide extreme sensitivity to quantify even small changes in receptor activity. With INDIGO’s cell-based reporter assays, the process is as easy as Thaw, Feed, Dose, and Read.
AhR Reporter Cells
AhR Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This proprietary cryopreservation process, enables long-term preservation of our unique reporter cells, so we can ship our cryopreserved reporter cells and assay reagents to you via overnight delivery, for your immediate use. Or, you can store the assay kits at -80°C. Once thawed, reporter cells are ready for immediate use so there is no need to take time on intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
AhR Luciferase Reporter
INDIGO’s AhR assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
AhR assay kits also available in: Human, Rat, Zebrafish
Aryl Hydrocarbon Receptor Assay Services
The primary application of INDIGO’s cell-based AhR assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of AhR. Service assays include a positive control reference compound and ‘vehicle’ control for every experiment. A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online “Request a Quote” form or contact an INDIGO Customer Service Representative to discuss your desired study parameters.
AhR Service Assays: Human, Rat, Zebrafish
Aryl Hydrocarbon Receptor Background
Although technically not a member of the Nuclear Receptor (NR) superfamily, the AhR shares many of the same attributes. The AhR is a member of the basic helix-loop-helix, Per-Arnt-Sim (bHLH-PAS) family of transcription factors and is responsible for the toxicologic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, often referred to as simply “dioxin”) and several other related polycyclic aromatic hydrocarbons. The basic mechanism of action of dioxin and related compounds has been extensively studied, in particular as it relates to regulation of cytochrome P450 1A1 (CYP1A1).
AhR is present in most cell types and in the unactivated state is cytosolic and exists in a complex with chaperone proteins such as heat shock protein 90 (Hsp90). Binding of TCDD and related molecules to AhR leads to nuclear translocation and heterodimerization with its partner protein ARNT, another bHLH-PAS family member. The AhR-ARNT heterodimer binds to specific cognate DNA sequence elements known as dioxin/xenobiotic response elements (DRE/XRE) present in the regulatory region of specific genes such as CYP1A1. Binding of the AhR:ARNT heterodimer to these elements, and subsequent recruitment of transcription coactivator complexes, leads to increased transcription of the specific gene, known as “target genes.” There is a battery of genes affected in this manner and targets include certain xenobiotic-metabolizing enzymes, such as CYP1A1,CYP1A2, CYP2B1, and UGT1A6. In addition, genes affected directly and indirectly by the TCDD/AhR-complex code for both inhibitory and stimulatory growth factors and their gene products affect cellular growth and differentiation leading to tumor promotion and carcinogenicity as well as other forms of toxicity.
INDIGO’s Aryl Hydrocarbon Receptor (AhR) Assay is a cell-based genetic reporter assay that utilizes the luciferase reporter gene Aryl Hydrocarbon Receptor Agonist and Antagonist Assays functionally linked to an AhR-responsive promoter. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in AhR activity.
The principle application of this reporter assay system is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against human AhR.
For more information on AhR, visit the Nuclear Receptor Resource.
Synonyms: Aryl Hydrocarbon Receptor, AhR, Ah Receptor, Dioxin Receptor
Aryl Hydrocarbon Receptor Assay Data
Agonist dose-response analyses of Human AhR.
Agonist analyses of Human AhR Reporter Cells were performed according to the protocol described in the assay technical manual, using the reference agonists MeBIO (provided), FICZ (6-Formylindolo(3,2-b)carbazole; Enzo), ITE (2-(1H-indole-3-ylcarbonyl)-4-thiazolecarboxylic methyl ester; Tocris), β-Napthoflavone (Sigma), Omeprazole and Pifthrin-a-hydrobromide (each from Tocris). Average relative light units (RLU) and corresponding standard deviation (SD) values were determined for each treatment concentration (n ≥ 6). Fold-activation (i.e., S/B) and Z’ values were calculated as described by Zhang, et al. (1999). Least squares fit non-linear regression and EC50 analyses were performed using GraphPad Prism software.
Antagonist dose-response analyses of Human AhR.
Antagonist analyses of Human AhR Reporter Cells were performed according to the protocol described in the assay technical manual, using the reference antagonists GNF351 (Calbiochem) and CH 223191 (Tocris).
Aryl Hydrocarbon Receptor Assay Research Areas
Artherosclerosis; Autoimmune Disease ; Cancer; Xenobiotic Metabolism; Bile Acid and Xenobiotic; Toxicology; Drug-Nutrient Interaction; Environmental Toxicology