Human E-Cadherin Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human E-Cadherin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human E-Cadherin in cell culture supernates, serum, plasma, urine, and saliva. It contains NS0-expressed recombinant human E-Cadherin/Fc Chimera and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human E-Cadherin 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 E-Cadherin.

Preparation and Storage

Background:  E-Cadherin

The cadherin superfamily comprises a large number of cell surface glycoproteins with one or more cadherin repeats, which are involved in Ca2⁺-dependent cell-cell adhesion. Classical cadherins contain five extracellular cadherin repeats and a conserved cytoplasmic region that interacts with the actin cytoskeleton via catenins. In addition to cell adhesion, cadherins also mediate morphogenesis, cytoskeletal organization, and cell migration. In addition to the classical cadherins, the cadherin superfamily also includes desmosomal cadherins, protocadherins, 7TM cadherins, FAT family cadherins, and T-cadherin.

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.