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Human AVPR1B Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays—
3 x-32 assays in-96 well format—
1 x-384 well format assays—
SIZE SKU
3 x-32 assays in-96 well format
1 x-384 well format assays
1 x-96 well format assays

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Arginine Vasopressin Receptor 1B (AVPR1B). INDIGO’s AVPR1B reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the AVPR1B. In addition to AVPR1B 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 AVPR1B. 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

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeGPCR
SpeciesHuman
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • Human AVPR1B Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Vasopressin, (ref. agonist; in PBS+0.1%BSA)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Activation of AVPR1B. Activation assays were performed using the reference polypeptides Vasopressin (provided), Felypressin, Terlipressin and Oxytocin. Luminescence was quantified and values of average (n = 3) relative light units (RLU), corresponding standard deviation (SD), Fold- Activation, and Z’ were calculated. The least-squares method of non-linear regression was used to plot Fold-Activation or RLU vs. Log10 [Compound, nM] and EC50 / IC50 values were determined using GraphPad Prism software.
Inhibition of AVPR1B. AVPR1B reporter cells were co-treated with an EC80 concentration of the activator vasopressin and varying concentrations of the AVPR1B specific inhibitor SSR149415. INDIGO’s Live Cell Multiplex (LCM) Assay confirmed that no treatment concentrations were cytotoxic (data not shown). Luminescence was quantified and values of average (n = 3) relative light units (RLU), corresponding standard deviation (SD), Fold- Activation, and Z’ were calculated. The least-squares method of non-linear regression was used to plot Fold-Activation or RLU vs. Log10 [Compound, nM] and EC50 / IC50 values were determined using GraphPad Prism software.

Target Background

The Arginine Vasopressin Receptor 1B (AVPR1B), also known as vasopressin 3 receptor, is a member of the family of G-Protein-coupled receptors (GPCR). There are three subtypes of vasopressin receptors identified in humans, which are AVPR1A, AVPR1B and AVPR2.

The natural ligand of AVPR1B is vasopressin, which is synthesized in the hypothalamus and secreted into the posterior pituitary. Like the closely similar neuropeptide, oxytocin, vasopressin is a nine amino acid neuropeptide that is classified as an antidiuretic hormone(ADH). Release of vasopressin is mediated by stimuli such as hemorrhage or dehydration. When vasopressin binds to AVPR1B signal transduction is mediated through Gaq/11 GTP binding proteins.

Like their respective ligands, vasopressin receptors and the oxytocin receptor (OXTR) are structurally similar (e.g., 85% between AVPR1A and OXTR)5. However, ligand selectivity for those receptors is different. The affinity of vasopressin for AVPR1A and AVPR1B is similar to that of the OXTR, whereas oxytocin has a significantly higher affinity for OXTR relative to the vasopressin receptors.

Although many peripheral tissues such as kidney, thymus, heart, lung, and spleen express AVPR1B, high expression of AVPR1B was detected in the anterior pituitary. The major role of AVPR1B in the brain is to stimulate production and secretion of adrenocorticotropin (ACTH) and cortisol . It has been suggested that AVPR1B is synergistically involved in the stress-induced activation of stress-reactive hypothalamic-pituitary-adrenal (HPA) axis with corticotropin-releasing hormone receptor-1 (CRHR1).

Due to its earlier discovery, AVPR1A has been studied more intensively than AVPR1B. However, AVPR1B is recognized as an important modulator of the endocrine stress response. Indeed, several knockout mice studies indicate that AVPR1B plays important roles in modulating such behaviors as social recognition memories and normal aggression.

Citations

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Arginine Vasopressin Receptor 1B (AVPR1B)

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