• Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Cell Lysates (25 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (13 uL)
  • Sensitivity
    16.6 pg/mL
  • Assay Range
    39.1 - 2,500 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant human Nectin-4.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC133, Quantikine Immunoassay Control Set 803 for Human Nectin-4 - Please Inquire
Product Summary
The Quantikine Human Nectin-4 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Nectin-4 in cell culture supernates, cell lysates, serum, plasma, and urine. It contains NS0-expressed recombinant human Nectin-4 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Nectin-4 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Nectin-4.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Cell Lysates, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean16342513651944741440
Standard Deviation5.5613.932.715.827.965.8
CV%3.43.32.48.15.94.6

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean23260318672455951801
Standard Deviation7.1937.740.118.931.982.3
CV%3.16.32.17.75.44.6

Recovery

The recovery of human Nectin-4 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 100 94-110
EDTA Plasma (n=4) 89 80-109
Heparin Plasma (n=4) 91 82-99
Serum (n=4) 90 80-101
Urine (n=4) 100 93-108
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Nectin-4 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Nectin-4 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Nectin-4

Nectins are type I transmembrane glycoproteins that are calcium-independent immunoglobulin (Ig)-like cell adhesion molecules (CAMs). Homology has led to their designation as poliovirus receptor-related (PRR) proteins, but nectins do not bind poliovirus. The extracellular domains of the nectin family form cis-homodimers (same cell), followed by homophilic or heterophilic trans-dimers (across cells) that organize intercellular junctions. Nectin-1, also known as CD111, PRR1 and herpes virus entry mediator C (HVEC) has a secreted splice variant. Nectin-1, Nectin-2 (CD112, PRR2, HVEB) and Nectin-3 (PRR3) are concentrated in adherens junctions and exist on neurons, endothelial cells, epithelial cells and fibroblasts.

Nectin-4 is a type I transmembrane glycoprotein belonging to the Nectin family of Ig superfamily proteins. It is both a homophilic and heterophilic (with Nectin-1) cell adhesion molecule that is expressed in the placenta, the embryo, and in breast carcinoma. A soluble form of Nectin-4 is generated from the membrane protein via the action of TACE/ADAM-17.

    • Long Name
      Poliovirus Receptor Related 4
    • Entrez Gene IDs
      81607 (Human); 71740 (Mouse);
    • Alternate Names
      EDSS1; Ig superfamily receptor LNIR; LNIR; LNIRnectin 4; Nectin4; Nectin-4; poliovirus receptor-related 4; poliovirus receptor-related protein 4; PRR4; PRR4DKFZp686K05193; PVRL4;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    The conjugate must remain at 2-8 °C during use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at 2-8 °C for 2 hours.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL cold Conjugate
    9.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    1 Citations: Showing 1 - 1

    1. Ectodomain Shedding of the Cell Adhesion Molecule Nectin-4 in Ovarian Cancer is Mediated by ADAM10 and ADAM17
      Authors: PC Buchanan, KL Boylan, B Walcheck, R Heinze, MA Geller, PA Argenta, AP Skubitz
      J. Biol. Chem, 2017;0(0):.
      Species: Human
      Sample Type: Serum
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 803 for Human Nectin-4

    Ctrl QC133
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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