Human VEGF-C Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human VEGF-C Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human VEGF-C in cell culture supernates, serum, plasma, and saliva. It contains NS0-expressed recombinant human VEGF-C (Cys156Ser substituted) and has been shown to accurately quantitate the recombinant factor. Results obtained using recombinant human VEGF-C (wild type) and natural human VEGF-C 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 VEGF-C.

Preparation and Storage

Background:  VEGF-C

Vascular endothelial growth factor C (VEGF-C) is a homodimeric ligand of VEGF R3/Flt-4 and is synthesized with N- and C-terminal propeptides. Fully processed VEGF-C containing only the VEGF homology domain can additionally bind and activate VEGF R2/KDR/Flk-1. VEGF-C interactions with VEGF R3 are critical for lymphangiogenesis. Both the ligand and receptor are usually co-expressed at sites of lymphatic vessel sprouting, in the embryo, and in various pathological conditions. Over-expression of VEGF-C in tumor cells induces tumoral lymphatic hyperplasia, neoangiogenesis, and vessel sprouting, resulting in enhanced lymph flow and metastasis to regional lymph nodes.

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.