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Human AVPR1A Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays
1 x-384 well format assays
SIZE SKU
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 1A (AVPR1A). INDIGO’s AVPR1A reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the AVPR1A. In addition to AVPR1A 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 AVPR1A. 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 AVPR1A 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 AVPR1A. 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 AVPR1A. AVPR1A reporter cells were co-treated with an EC80 concentration of the activator vasopressin and varying concentrations of the AVPR1A inhibitors, SR 49059, Conivaptan and TC OT39. 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 1A (AVPR1A) 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 AVPR1A 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 AVPR1A 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). However, ligand selectivity for those receptors is different. The affinity of vasopressin for AVPR1A and AVPR1B is same as OXTR, whereas oxytocin has a significantly higher affinity for OXTR than for the vasopressin receptors.

The physiological roles induced by AVPR1A vary in both the peripheral and central system. For example, AVPR1A is highly distributed on vascular smooth muscle and plays a role as a vasoconstrictor to maintain water balance in our body. Expression of AVPR1A is also detected in the liver, kidney and the uterus. The distribution of AVPR1A is not only limited to the peripheral tissues but also expressed in the brain. AVPR1A has been implicated to play a role in social cognition and behavior in humans.

Much effort has been made to investigate the roles of AVPR1A and to identify bioactive peptide analogues and small molecule ligands. Many peptide analogues are utilized as agonists to induce antidiuretic or vasoconstricting activities. In contrast, small molecules are better therapeutic agents as antagonists due to relatively improved bioavailability and stability. Several clinical approaches using AVPR1A antagonists have been shown to improve social communications and social sensitivity in Autism.

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Arginine Vasopressin Receptor 1A (AVPR1A)

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