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Human Nectin-4 ELISA Kit - Quantikine

Catalog #: DNEC40
2 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DNEC40
Control Products Available
QC133: Quantikine Immunoassay Control Set 1063 for Human Nectin-4
Human Nectin-4 ELISA Serum/Plasma Standard Curve
3 Images
Product Details
Procedure
Citations (2)
FAQs
Supplemental Products
Reviews
Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human Nectin-4 Quantikine ELISA Kit Summary

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.

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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 163 425 1365 194 474 1440
Standard Deviation 5.56 13.9 32.7 15.8 27.9 65.8
CV% 3.4 3.3 2.4 8.1 5.9 4.6

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 232 603 1867 245 595 1801
Standard Deviation 7.19 37.7 40.1 18.9 31.9 82.3
CV% 3.1 6.3 2.1 7.7 5.4 4.6

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 ELISA Linearity

Scientific Data

N/A Human Nectin-4 ELISA Serum/Plasma Standard Curve View Larger

Human Nectin-4 ELISA Serum/Plasma Standard Curve

N/A Human Nectin-4 ELISA Cell Culture Supernate/Cell Lysate/Urine Standard Curve View Larger

Human Nectin-4 ELISA Cell Culture Supernate/Cell Lysate/Urine Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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); 498281 (Rat); 102127936 (Cynomolgus Monkey)
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
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL cold Conjugate
  6.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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 for Human Nectin-4 Quantikine ELISA Kit

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.

2 Citations: Showing 1 - 2
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  1. Genetic variants in NECTIN4 encoding an adhesion molecule are associated with continued opioid use
    Authors: CP Fang, TH Liu, RH Chung, HH Tsou, HW Kuo, SC Wang, CC Liu, SC Liu, ACH Chen, YL Liu
    PLoS ONE, 2020-06-18;15(6):e0234549.
    Species: Human
    Sample Types: Plasma
  2. 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, 2017-02-23;0(0):.
    Species: Human
    Sample Types: Serum

FAQs

  1. Does Human Nectin-4 Quantikine ELISA kit detect the soluble and/or total protein?

    • This kit detects both soluble and transmembrane Nectin-4, confirmed by sample measurements in cell culture supernatants and the corresponding cell lysates.

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