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Human ADRB3 Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays
3 x-32 assays in-96 well format
SIZE SKU
3 x-32 assays in-96 well format
1 x-96 well format assays

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the the Human Adrenoreceptor Beta 3 (ADRB3). INDIGO’s ADRB1 reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the ADRB3. In addition to ADRB1 Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. 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 ADRB3. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.

Features

  • Ready to Use Upon Receipt

  • Includes All Needed Components
  • Contains Transfected Reporter Cells
  • Eliminates Cell Licensing Fees
  • Clear, Reproducible Results
  • Consistent Results Lot to Lot

Product Specifications

Target TypeGPCR
SpeciesHuman
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • Human ADRB3 Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Norepinephrine, (ref. agonist; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Activation of ADRB3. Reporter cells were treated with the reference activators Isoproterenol (provided), CGP 12177, Epinephrine, Norepinephrine, and Mirabegron. The absence of signal in Isoproterenol treated ‘Mock’ cells (which contain the CRE-Luc reporter vector, but do not express ADRB3) confirms that the observed ligand-dependent response is specific to ADRB3 activation.
Inhibition of ADRB3. Reporter cells were co-treated with an EC80 concentration of the reference activator Isoproterenol and varying concentrations of the b3 adrenoceptors selective inhibitor L-748,337, and the non-selective b-adrenoceptor inhibitors Timolol and Propranolol, and the selective b2-adrenoceptor inhibitor ICI 118551. INDIGO’s Live Cell Multiplex (LCM) Assay confirmed that no treatment concentrations were cytotoxic (data not shown).

Target Background

The adrenoceptors (a.k.a. adrenergic receptors) mediate the action of the sympathetic nervous system and are activated in response to “fight-or-flight” signals. They are divided into three types, adrenoceptor α1-, α2-, and β. Each type is further composed of three subtypes resulting in 9 different types (α1A, α1B, α1D, α2A, α2B, α2C, β1, β2, and β3). Adrenoceptors belong to the G-Protein-coupled receptor (GPCR) family. They all display the characteristic seven transmembrane helices, the extracellular loops which contribute to ligand binding, and the intracellular carboxy tail that associates with trimeric G proteins. All nine types of adrenoceptors are activated by the same endogenous catecholamines (epinephrine and norepinephrine); however, the specificity of their responses depends on the G-proteins and effector systems they associate with in a tissue and time specific manner.

Adrenoceptor Beta 3 (ADRB3) was the last discovered member of the b-adrenoceptor sub-family and is the least studied. However, its utility as a therapeutic target has been validated by the approval of the ADRB3 agonists Mirabegron and Vibegron, which are used to treat overactive bladder syndrome. An additional feature that makes it a potential target for chronic conditions is that it lacks sites for phosphorylation by protein kinase A and b-adrenoceptor kinase, which are present on the b1 and b2-adrenoceptors, making it potentially resistant to desensitization. ADRB3 is of additional interest as a target for the treatment of metabolic disorders, due to its role in lipolysis and thermogenesis and its expression in white and beige human adipocytes. Further, targeting ADRB3 may be useful for the treatment of cancer, since it has been demonstrated that ADRB3 is over-expressed in the tumor microenvironment, and has also been implicated in other processes including angiogenesis and cancer progression. Its involvement and expression in a variety of physiological processes and cell types, respectively, makes it a promising target in the treatment of multiple diseases.

Citations

Also available as a service

Adrenoreceptor Beta 3 (ADRB3)

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