Human uPAR Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human uPAR Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human uPAR in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human uPAR and antibodies raised against the recombinant factor. It has been shown to accurately quantitate the recombinant factor. Results obtained using natural human uPAR 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 of natural human uPAR.

Preparation and Storage

Background: uPAR

The urokinase-type Plasminogen Activator Receptor (uPAR) is a GPI-anchored protein that binds to both the inactive and active forms of uPA. This interaction serves to localize and enhance the proteolytic activity of uPA, leading to increased conversion of Plasminogen to Plasmin. uPA also triggers uPAR-mediated signal transduction, activation of protein tyrosine kinases, gene expression, cell adhesion, and chemotaxis. uPAR can additionally regulate cell adhesion by direct binding to Vitronectin and preventing Integrins from binding to their ligands.

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.