Human Nectin-4 Quantikine ELISA Kit Summary

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.

Preparation and Storage

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.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all 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、Add 50 μL of Assay Diluent to each well.

4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7、Aspirate and wash 5 times.

8、Add 100 μL Substrate Solution to each well.

9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.