Human Nectin-4 Quantikine ELISA Kit

R&D Systems | Catalog # DNEC40

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Cell Lysates (25 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Urine (13 µL)

Sensitivity

16.6 pg/mL

Assay Range

39.1-2500 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
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Product Summary for Human Nectin-4 Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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

EDTA Plasma, Heparin Plasma, Serum

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 for Human Nectin-4 Quantikine ELISA Kit

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 Images for Human Nectin-4 Quantikine ELISA Kit

Human Nectin-4 ELISA Serum/Plasma Standard Curve

Human Nectin-4 ELISA Serum/Plasma Standard Curve

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

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

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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

Alternate Names

LNIR, Nectin4, PRR4, PVRL4

Entrez Gene IDs

81607 (Human); 71740 (Mouse); 498281 (Rat); 102127936 (Cynomolgus Monkey)

Gene Symbol

NECTIN4

Additional Nectin-4 Products

Product Documents for Human Nectin-4 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Nectin-4 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human Nectin-4 Quantikine ELISA Kit

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Protocols

View specific protocols for Human Nectin-4 Quantikine ELISA Kit (DNEC40):

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.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human Nectin-4 Quantikine ELISA Kit

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  • Q: Does Human Nectin-4 Quantikine ELISA kit detect the soluble and/or total protein?

    A: 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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