Zebrafish Aryl Hydrocarbon Receptor (zAhR)
|Product Family||Product Number||Product Description||Technical Manual|
|Z06001-32||Zebrafish AhR Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|Z06001||Zebrafish AhR Reporter Assay System, 1 x 96-well format assays||Technical Manual|
Kits are offered in different assay formats to accommodate researchers’ needs: 3x 32 and 1x 96, assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications. Assay systems are all inclusive, providing reporter cells, optimized growth media, media for diluting test compounds, a positive-control agonist, luciferase detection reagent, a white assay plate, a detailed protocol, and a protocol quick guide. All kits are shipped on dry ice.
zAhR Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This cryo-preservation method yields high cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay plates. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.
INDIGO’s zAhR assay kit is an all-inclusive system. In addition to reporter cells, this kit provides two optimized media for use during cell culture and in diluting the user's test samples, the AhR reference agonist MeBIO, Luciferase Detection Reagent, and a cell culture-ready assay plate.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
INDIGO's Zebrafish Aryl Hydrocarbon Receptor (zfAhR) Reporter Cells are constructed in zebrafish ZF4 cells, a cell line derived from Brachydanio rerio embryos. This zebrafish cell line has been modified to contain the luciferase reporter gene functionally linked to tandem DRE/XRE genetic response elements and a minimal promoter sequence. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against the native zebrafish AhR.
While technically not a nuclear receptor, the AhR is mechanistically and functionally similar to members of that super-family, being both a receptor and a ligand-activated transcription factor. More formally, the AhR is a member of the basic helix-loop-helix, Per-Arnt-Sim family of transcription factors. As its name implies, the Aryl Hydrocarbon Receptor (AhR) is a xenobiotic-sensing receptor. It is responsible for the adverse toxicologic effects elicited by various polycyclic aromatic hydrocarbon environmental and industrial pollutants, perhaps the most infamous being 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).
The AhR is present in the cytosol where, in the non-active state, it is in a complex with chaperone proteins such as Hsp90. Binding of a polycyclic aromatic hydrocarbon to AhR leads to nuclear translocation and association with its partner protein ARNT. The AhRARNT hetero-dimer binds to specific cognate DNA sequence elements known as dioxin/xenobiotic response elements (DRE/XRE) present in the regulatory region of a variety of target genes. Binding of AhR:ARNT to these elements, and subsequent recruitment of transcription co-activator complexes, induces the transcription of a battery of target genes, including several cytochrome P450 genes. Other target genes of the TCDD/AhR-complex encode inhibitory or stimulatory growth factors that mediate cellular growth and differentiation.
For more information on AhR, visit the Nuclear Receptor Resource.
The primary application of INDIGO’s cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given receptor. 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.
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