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.
Key Product Details
Assay Length
4.25 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Citrate Plasma (50 µL)
Sensitivity
40 pg/mL
Assay Range
125-8000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Citrate Plasma)
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Product Summary for Human HGF Quantikine ELISA Kit
The Quantikine Human HGF Immunoassay is a 4.25 or 4.5 hour solid phase ELISA that is designed to measure HGF levels in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human pro-HGF and antibodies raised against the recombinant factor and has been shown to quantitate recombinant human HGF accurately. Results obtained using natural 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 HGF.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human HGF
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence
of human HGF in this assay. No medical histories were available for the donors used in this
study.
Cell Culture Supernates - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal calf serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 1 and 5 days. Aliquots of the cell culture supernates were removed and assayed for levels of human HGF.
ND=Non-detectable
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=60) | 1257 | 671-1992 | 289 |
| EDTA plasma (n=60) | 787 | 469-113 | 136 |
| Citrate plasma (n=71) | 440 | 251-742 | 102 |
Cell Culture Supernates - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal calf serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 1 and 5 days. Aliquots of the cell culture supernates were removed and assayed for levels of human HGF.
| Condition | Day 1 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | ND | 548 |
| Stimulated | 152 | 145 |
Precision
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.
Cell Culture Supernates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 425 | 991 | 3869 | 409 | 1058 | 4058 |
| Standard Deviation | 30.2 | 43.2 | 234 | 29.4 | 75.1 | 264 |
| CV% | 7.1 | 4.4 | 6.0 | 7.2 | 7.1 | 6.5 |
Citrate Plasma, EDTA Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 339 | 999 | 3759 | 363 | 967 | 3790 |
| Standard Deviation | 23.7 | 56.2 | 154 | 25.8 | 81.2 | 205 |
| CV% | 7.0 | 5.6 | 4.1 | 7.1 | 8.4 | 5.4 |
Recovery for Human HGF Quantikine ELISA Kit
The recovery of HGF spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 92 | 86-101 |
| Citrate Plasma (n=5) | 105 | 99-114 |
| EDTA Plasma (n=5) | 100 | 85-111 |
| Serum (n=5) | 99 | 91-108 |
Linearity
To assess the linearity of the assay, five samples of each sample type were spiked with high concentrations of HGF and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human HGF Quantikine ELISA Kit
Human HGF ELISA Standard Curve
Human HGF ELISA Cell Culture Supernate Standard Curve
Human HGF ELISA Serum/Plasma Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: HGF
Long Name
Hepatocyte Growth Factor
Alternate Names
DFNB39, F-TCF, Hepatopoietin A, HGFB, HPTA, SF
Entrez Gene IDs
Gene Symbol
HGF
Additional HGF Products
Product Documents for Human HGF Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human HGF Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human HGF Quantikine ELISA Kit
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Protocols
View specific protocols for Human HGF Quantikine ELISA Kit (DHG00):
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.
- Add 150 µL of Assay Diluent to each well.
- 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 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well.
- For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1.75 hours. - Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
150 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
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