Natural and recombinant HGF. Recombinant mouse HGF single chain precursor cross-reacts 100% in this assay.
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Mouse/Rat HGF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse or rat HGF in cell culture supernates, tissue homogenates, serum, and platelet-poor plasma. It contains NS0-expressed recombinant mouse HGF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural HGF showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural HGF.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
The recovery of HGF spiked into various matrices was evaluated.
Average % Recovery
Mouse Cell Culture Supernates (n=4)
Mouse Platelet-poor EDTA Plasma (n=4)
Mouse Serum (n=4)
Mouse Tissue Homogenates (n=3)
Rat Cell Culture Supernates (n=4)
Rat Platelet-poor EDTA Plasma (n=4)
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse/rat HGF were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
Refer to the product for 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.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 5 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
100 µL Stop Solution
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.
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