HGF ELISA Kit (Hepatocyte Growth Factor (Hepapoietin A, Scatter Factor))

Details for Product HGF ELISA Kit No. ABIN1979644, Supplier: Log in to see
Antigen
  • DFNB39
  • F-TCF
  • HGFB
  • HPTA
  • SF
  • HGF
  • C230052L06Rik
  • HGF/SF
  • NK1
  • NK2
  • SF/HGF
  • hepatocyte growth factor
  • HGF
  • Hgf
Reactivity
Human
Alternatives
Kits with alternative reactivity to:
38
24
24
16
11
11
6
3
2
1
1
1
1
1
Method Type
Sandwich ELISA
Detection Range
3-2000 pg/mL
Minimum Detection Limit
3 pg/mL
Application
ELISA
Options
Supplier
Log in to see
Supplier Product No.
Log in to see
Purpose Human HGF ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Plasma, Cell Culture Supernatant, Serum
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
Sensitivity < 3 pg/mL
Characteristics
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Material not included
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Plasmids, Primers & others Plasmids, Primers & others HGF products on genomics-online (e.g. as negative or positive controls)
Antigen
Alternative Name HGF (HGF ELISA Kit Abstract)
Background The Human HGF ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human HGF in serum, plasma (EDTA and citrate), cell culture supernatants and urine. This assay employs an antibody specific for human HGF coated on a 96-well plate. Standards and samples are pipetted into the wells and HGF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human HGF antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of HGF bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
Gene ID 3082
UniProt P14210
Pathways RTK Signaling, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Synaptic Membrane, Signaling of Hepatocyte Growth Factor Receptor
Application Notes Recommended Dilution for serum and plasma samples2 fold
Sample Volume 100 μL
Plate Pre-coated
Protocol
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Reagent Preparation
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
    2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
    3. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
    4. Preparation of standard: Briefly spin the vial of Item C and then add 600 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 2,000 pg/mL standard. Dissolve the powder thoroughly by a gentle mix. Pipette 260 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). Standard + 600 µL 130 µL 130 µL 130 µL 130 µL 130 µL 130myl 2000 666.7 222.2 74.07 24.7 8.23 2.74 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
    5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
    6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
    7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 400-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 35 µL of HRP-Streptavidin concentrate into a tube with 14 ml 1x Assay Diluent B to prepare a final 400 fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
Assay Procedure
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
    2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
    3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
    4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
    5. Discard the solution. Repeat the wash as in step
    6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
    7. Discard the solution. Repeat the wash as in step
    8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
    9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Human HGF concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10 Assay Diluent B Human HGF concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of HGF is typically less than 3 pg/mL.
Recovery: Recovery was determined by spiking various levels of human HGF into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 108.28 96-120 Plasma 107.56 95-119 Cell culture media 98.35 89-108
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 95 94 97 Range ( %) 91-103 89-105 89-106 1:4 Average % of Expected 98 96 93 Range ( %) 90-106 88-105 91-108
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Assay Precision Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Restrictions For Research Use only
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date 6 months
Supplier Images
ELISA image for Hepatocyte Growth Factor (Hepapoietin A, Scatter Factor) (HGF) ELISA Kit (ABIN1979644) Hepatocyte Growth Factor (Hepapoietin A, Scatter Factor) (HGF) ELISA Kit
Product cited in: Nowak, Siemińska, Karpe, Marek, Kos-Kudła, Kajdaniuk: "Serum concentrations of HGF and IL-8 in patients with active Graves' orbitopathy before and after methylprednisolone therapy." in: Journal of endocrinological investigation, Vol. 39, Issue 1, pp. 63-72, 2016 (PubMed).

Koob, Lim, Zabek, Massee: "Cytokines in single layer amnion allografts compared to multilayer amnion/chorion allografts for wound healing." in: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 103, Issue 5, pp. 1133-40, 2015 (PubMed).

Drebert, Bracke, Beck: "Glucocorticoids and the non-steroidal selective glucocorticoid receptor modulator, compound A, differentially affect colon cancer-derived myofibroblasts." in: The Journal of steroid biochemistry and molecular biology, Vol. 149, pp. 92-105, 2015 (PubMed).

Jung, Han, Kim, Kim, Lee, Seo, Youn, Lee: "Stimulatory effect of HGF-overexpressing adipose tissue-derived mesenchymal stem cells on thymus regeneration in a rat thymus involution model." in: Cell biology international, 2014 (PubMed).

Stewart, Ramani, Sanderson: "Shed Syndecan-1 Translocates to the Nucleus of Cells Delivering Growth Factors and Inhibiting Histone Acetylation: A Novel Mechanism of Tumor-Host Crosstalk." in: The Journal of biological chemistry, 2014 (PubMed).

Koob, Lim, Massee, Zabek, Rennert, Gurtner, Li: "Angiogenic properties of dehydrated human amnion/chorion allografts: therapeutic potential for soft tissue repair and regeneration." in: Vascular cell, Vol. 6, pp. 10, 2014 (PubMed).

Fernández-Yunquera, Ripoll, Bañares, Puerto, Rincón, Yepes, Catalina, Salcedo: "Everolimus immunosuppression reduces the serum expression of fibrosis markers in liver transplant recipients." in: World journal of transplantation, Vol. 4, Issue 2, pp. 133-40, 2014 (PubMed).

Lee, Park, Jeon, Kim, Chang: "Potentiated therapeutic angiogenesis by primed human mesenchymal stem cells in a mouse model of hindlimb ischemia." in: Regenerative medicine, Vol. 8, Issue 3, pp. 283-93, 2013

Khan, Sharma, Kumar, Husain, Sharma: "Interrelationship and expression profiling of cyclooxygenase and angiogenic factors in Indian patients with multiple myeloma." in: Annals of hematology, Vol. 92, Issue 1, pp. 101-9, 2013 (PubMed).

Lee, Park, Jeon, Kim, Chang: "Potentiated therapeutic angiogenesis by primed human mesenchymal stem cells in a mouse model of hindlimb ischemia." in: Regenerative medicine, Vol. 8, Issue 3, pp. 283-93, 2013 (PubMed).

Najar, Raicevic, Fayyad-Kazan, Kazan, De Bruyn, Bron, Toungouz, Lagneaux: "Immune-related antigens, surface molecules and regulatory factors in human-derived mesenchymal stromal cells: the expression and impact of inflammatory priming." in: Stem cell reviews, Vol. 8, Issue 4, pp. 1188-98, 2012 (PubMed).

Nazarov, Lee, Soupene, Etemad, Knapik, Green, Bashkirova, Fang, Matthay, Kuypers, Serikov: "Multipotent stromal stem cells from human placenta demonstrate high therapeutic potential." in: Stem cells translational medicine, Vol. 1, Issue 5, pp. 359-72, 2012 (PubMed).

Burdon, Macgregor, Bykhovskaya, Javadiyan, Li, Laurie, Muszynska, Lindsay, Lechner, Haritunians, Henders, Dash, Siscovick, Anand, Aldave, Coster, Szczotka-Flynn, Mills, Iyengar, Taylor, Phillips et al.: "Association of polymorphisms in the hepatocyte growth factor gene promoter with keratoconus. ..." in: Investigative ophthalmology & visual science, Vol. 52, Issue 11, pp. 8514-9, 2012 (PubMed). (Sample species: Human). Further details: Sample: Serum (Genotypes of tag SNPs around HGF)

Tyan, Kuo, Huang, Pan, Shew, Chang, Lee, Lee: "Breast cancer cells induce cancer-associated fibroblasts to secrete hepatocyte growth factor to enhance breast tumorigenesis." in: PLoS ONE, Vol. 6, Issue 1, pp. e15313, 2011 (PubMed). (Sample species: Human). Further details: Sample: Conditioned Media

Shintani, Aoki, Nishihara, Ohno, Furusho, Hiromatsu, Akashi, Imaizumi, Aoyagi: "Hepatocyte growth factor promotes an anti-inflammatory cytokine profile in human abdominal aortic aneurysm tissue." in: Atherosclerosis, Vol. 216, Issue 2, pp. 307-12, 2011 (PubMed). (Sample species: Human). Further details: Sample: Conditioned Media

Raicevic, Najar, Stamatopoulos, De Bruyn, Meuleman, Bron, Toungouz, Lagneaux: "The source of human mesenchymal stromal cells influences their TLR profile as well as their functional properties." in: Cellular immunology, Vol. 270, Issue 2, pp. 207-16, 2011 (PubMed). (Sample species: Human). Further details: Sample: Conditioned Media (MSC)

Jalili, Shirvaikar, Marquez-Curtis, Turner, Janowska-Wieczorek: "The HGF/c-Met axis synergizes with G-CSF in the mobilization of hematopoietic stem/progenitor cells." in: Stem cells and development, Vol. 19, Issue 8, pp. 1143-51, 2010 (PubMed).

Did you look for something else?