Human HGF Quantikine ELISA Kit Summary

Product Summary

The Quantikine^® Human HGF Immunoassay is a 4.0 hour solid phase ELISA that is designed to measure human HGF levels in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human HGF and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human HGF. Results obtained using natural human HGF 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 natural human HGF.

Preparation and Storage

Background: HGF

HGF (Hepatocyte Growth Factor, Scatter Factor) induces the proliferation and migration of epithelial cells, hepatocytes, chondrocytes, keratinocytes, melanocytes, endothelial cells, and many tumor cells. During organogenesis and tissue repair, HGF promotes epithelial/endothelial morphogenesis by inducing cell scattering and branching tubulogenesis. It also supports insulin production by pancreatic beta cells, neuronal survival, and immune tolerance. HGF is secreted as a propeptide that is activated by uPA or HGF Activator at sites of tissue damage. Its signaling through the receptor HGF R/c-MET is enhanced by its prior binding to heparan sulfate proteoglycans. The serum levels of HGF are elevated in a wide range of pathologies including liver damage, acute kidney failure, myocardial infarction, type 1 diabetes, obesity, and cancer, as well as in the synovial fluid of rheumatoid arthritis patients.

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.