Human HGF DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human HGF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
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
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human HGF DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
29
Citations: Showing 1 - 10
Filter your results:
Filter by:
-
Function of hepatocyte spheroids in bioactive microcapsules is enhanced by endogenous and exogenous hepatocyte growth factor
Authors: Gwon, K;Choi, D;de Hoyos-Vega, JM;Baskaran, H;Gonzalez-Suarez, AM;Lee, S;Hong, HJ;Nguyen, KM;Dharmesh, E;Sugahara, G;Ishida, Y;Saito, T;Stybayeva, G;Revzin, A;
Bioactive materials
Species: Human
Sample Types: Microsphere Precipitate
-
Guiding Hepatic Differentiation of Pluripotent Stem Cells Using 3D Microfluidic Co-Cultures with Human Hepatocytes
Authors: Fattahi, P;de Hoyos-Vega, JM;Choi, JH;Duffy, CD;Gonzalez-Suarez, AM;Ishida, Y;Nguyen, KM;Gwon, K;Peterson, QP;Saito, T;Stybayeva, G;Revzin, A;
Cells
Species: Human
Sample Types: Cell Culture Supernates
-
Pro-cancerogenic effects of spontaneous and drug-induced senescence of ovarian cancer cells in vitro and in vivo: a comparative analysis
Authors: S Rutecki, P Szulc, M Paku?a, P Uruski, A Radziemski, E Naumowicz, R Moszy?ski, A Tykarski, J Miku?a-Pie, K Ksi??ek
Journal of ovarian research, 2022-07-26;15(1):87.
Species: Human
Sample Types: Cell Culture Supernates
-
Delivery of affordable and scalable encapsulated allogenic/autologous mesenchymal stem cells in coagulated platelet poor plasma for dental pulp regeneration
Authors: I Angelopoul, C Trigo, MI Ortuzar, J Cuenca, C Brizuela, M Khoury
Scientific Reports, 2022-01-10;12(1):435.
Species: Human
Sample Types: Cell Culture Supernates
-
Efficient blockade of locally reciprocated tumor-macrophage signaling using a TAM-avid nanotherapy
Authors: SJ Wang, R Li, TSC Ng, G Luthria, MJ Oudin, M Prytyskach, RH Kohler, R Weissleder, DA Lauffenbur, MA Miller
Sci Adv, 2020-05-22;6(21):eaaz8521.
Species: Human
Sample Types: Cell Lysates
-
Mesenchymal Stem and Stromal Cells Harness Macrophage-Derived Amphiregulin to Maintain Tissue Homeostasis
Authors: JH Ko, HJ Kim, HJ Jeong, HJ Lee, JY Oh
Cell Rep, 2020-03-17;30(11):3806-3820.e6.
Species: Human
Sample Types: Cell Culture Supernates
-
Are the Immune Properties of Mesenchymal Stem Cells from Wharton's Jelly Maintained during Chondrogenic Differentiation?
Authors: C Voisin, G Cauchois, L Reppel, C Laroye, L Louarn, C Schenowitz, P Sonon, I Poras, V Wang, E D Carosell, N Benkirane-, P Moreau, N Rouas-Frei, D Bensoussan, C Huselstein
J Clin Med, 2020-02-04;9(2):.
Species: Human
Sample Types: Cell Culture Supernates
-
A Unique Pattern of Mesothelial-Mesenchymal Transition Induced in the Normal Peritoneal Mesothelium by High-Grade Serous Ovarian Cancer
Authors: M Paku?a, P Uruski, A Niklas, A Wo?niak, D Szpurek, A Tykarski, J Miku?a-Pie, K Ksi??ek
Cancers (Basel), 2019-05-13;11(5):.
Species: Human
Sample Types: Cell Culture Supernates
-
Hepatic blood flow by perfusion computed tomography as an imaging biomarker for patients with gastric cancer
Authors: K Shuto, M Mori, C Kosugi, K Narushima, S Nakabayash, T Fujisiro, A Sato, K Hayano, H Shimizu, K Koda
Oncol Lett, 2019-01-25;17(3):3267-3276.
Species: Human
Sample Types: Serum
-
Four types of human platelet lysate, including one virally inactivated by solvent-detergent, can be used to propagate Wharton Jelly mesenchymal stromal cells
Authors: MS Chen, TJ Wang, HC Lin, B Thierry
New biotechnology, 2018-11-20;0(0):.
Species: Human
Sample Types: Platelet Lysates
-
TGF beta inhibits HGF, FGF7, and FGF10 expression in normal and IPF lung fibroblasts
Authors: KA Correll, KE Edeen, EF Redente, RL Zemans, BL Edelman, T Danhorn, D Curran-Eve, A Mikels-Vig, RJ Mason
Physiol Rep, 2018-08-01;6(16):e13794.
Species: Human
Sample Types: Cell Culture Supernates
-
Comparative Analysis of Different Platelet Lysates and Platelet Rich Preparations to Stimulate Tendon Cell Biology: An In Vitro Study
Authors: F Klatte-Sch, T Schmidt, M Uckert, S Scheffler, U Kalus, M Rojewski, H Schrezenme, A Pruss, B Wildemann
Int J Mol Sci, 2018-01-10;19(1):.
Species: Human
Sample Types: Plasma
-
The Proangiogenic Capabilities of Malignant Ascites Generated by Aggressive Ovarian Tumors
Authors: J Miku?a-Pie, P Uruski, S Szubert, K Maksin, R Moszy?ski, D Szpurek, A Wo?niak, S Sajdak, A Tykarski, K Ksi??ek
Biomed Res Int, 2017-09-20;2017(0):2592496.
Species: Human
Sample Types: Cell Culture Supernates
-
Reduced microRNA-503 expression augments lung fibroblast VEGF production in chronic obstructive pulmonary disease
Authors: J Ikari, AJ Nelson, J Obaid, A Giron-Mart, K Ikari, F Makino, S Iwasawa, Y Gunji, M Farid, X Wang, H Basma, D Demeo, C Feghali-Bo, O Holz, K Rabe, X Liu, SI Rennard
PLoS ONE, 2017-09-07;12(9):e0184039.
Species: Human
Sample Types: Cell Culture Supernates
-
A Collagen Cardiac Patch Incorporating Alginate Microparticles Permits the Controlled Release of HGF and IGF-1 to Enhance Cardiac Stem Cell Migration and Proliferation
Authors: Hugh S O'Neill
J Tissue Eng Regen Med, 2017-06-19;0(0):.
-
mTORC1-independent autophagy regulates receptor tyrosine kinase phosphorylation in colorectal cancer cells via an mTORC2-mediated mechanism
Authors: A Lampada, J O'Prey, G Szabadkai, KM Ryan, D Hochhauser, P Salomoni
Cell Death Differ., 2017-05-05;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
-
Effect of 2-octylcyanoacrylate on placenta derived mesenchymal stromal cells on extracellular matrix
Authors: YJ Chen, L Lankford, S Kabagambe, Z Saenz, P Kumar, D Farmer, A Wang
Placenta, 2017-04-02;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
-
Secretory activity of subcutaneous abdominal adipose tissue in male patients with rheumatoid arthritis and osteoarthritis - association with clinical and laboratory data
Authors: Ewa Kontny
Reumatologia, 2016-11-28;54(5):227-235.
Species: Human
Sample Types: Tissue Extracts
-
Characterization of In Vitro Engineered Human Adipose Tissues: Relevant Adipokine Secretion and Impact of TNF-alpha.
Authors: Aubin K, Safoine M, Proulx M, Audet-Casgrain M, Côté J, Tetu F, Roy A, Fradette J
PLoS ONE, 2015-09-14;10(9):e0137612.
Species: Human
Sample Types: Cell Culture Supernates
-
Creating capillary networks within human engineered tissues: Impact of adipocytes and their secretory products
Authors: Kim Aubin, Caroline Vincent, Maryse Proulx, Dominique Mayrand, Julie Fradette
Acta Biomaterialia
Species: Human
Sample Types: Cell Culture Supernates
-
Aqueous two-phase system patterning of detection antibody solutions for cross-reaction-free multiplex ELISA.
Authors: Frampton, John P, White, Joshua B, Simon, Arlyne B, Tsuei, Michael, Paczesny, Sophie, Takayama, Shuichi
Sci Rep, 2014-05-02;4(0):4878.
Species: Human
Sample Types: Plasma
-
Fibroblasts derived from human pluripotent stem cells activate angiogenic responses in vitro and in vivo.
Authors: Shamis Y, Silva E, Hewitt K, Brudno Y, Levenberg S, Mooney D, Garlick J
PLoS ONE, 2013-12-30;8(12):e83755.
Species: Human
Sample Types: Cell Culture Supernates
-
Transplantation of human adipose tissue-derived stem cells delays clinical onset and prolongs life span in ALS mouse model.
Authors: Kim, Kwang S, Lee, Hong J, An, Jin, Kim, Yun B, Ra, Jung Cha, Lim, Inja, Kim, Seung U
Cell Transplant, 2013-09-18;23(12):1585-97.
Species: Human
Sample Types: Cell Culture Supernates
-
Heparanase Plays a Dual Role in Driving Hepatocyte Growth Factor (HGF) Signaling by Enhancing HGF Expression and Activity.
Authors: Ramani VC, Yang Y, Ren Y
J. Biol. Chem., 2010-12-03;286(8):6490-9.
Species: Human
Sample Types: Cell Culture Supernates
-
Angiogenesis and lymphangiogenesis are downregulated in primary breast cancer.
Authors: Boneberg EM, Legler DF, Hoefer MM, Ohlschlegel C, Steininger H, Fuzesi L, Beer GM, Dupont-Lampert V, Otto F, Senn HJ, Furstenberger G
Br. J. Cancer, 2009-08-18;101(4):605-14.
Species: Human
Sample Types: Serum
-
Regulation of hepatocyte growth factor secretion by fibroblasts in patients with acute lung injury.
Authors: Quesnel C, Marchand-Adam S, Fabre A, Marchal-Somme J, Philip I, Lasocki S, Lecon V, Crestani B, Dehoux M
Am. J. Physiol. Lung Cell Mol. Physiol., 2007-12-07;294(2):L334-43.
Species: Human
Sample Types: BALF
-
Influence of microvascular endothelial cells on transcriptional regulation of proximal tubular epithelial cells.
Authors: Aydin S, Signorelli S, Lechleitner T, Joannidis M, Pleban C, Perco P, Pfaller W, Jennings P
Am. J. Physiol., Cell Physiol., 2007-12-05;294(2):C543-54.
Species: Human
Sample Types: Cell Culture Supernates
-
Bronchial epithelial cell growth regulation in fibroblast cocultures: the role of hepatocyte growth factor.
Authors: Skibinski G, Elborn JS, Ennis M
Am. J. Physiol. Lung Cell Mol. Physiol., 2007-03-23;293(1):L69-76.
Species: Human
Sample Types: Cell Culture Supernates
-
Structural basis of hepatocyte growth factor/scatter factor and MET signalling.
Authors: Gherardi E, Sandin S, Petoukhov MV, Finch J, Youles ME, Ofverstedt LG, Miguel RN, Blundell TL, Vande Woude GF, Skoglund U, Svergun DI
Proc. Natl. Acad. Sci. U.S.A., 2006-03-06;103(11):4046-51.
Species: Human
Sample Types: Cell Culture Supernates
FAQs
No product specific FAQs exist for this product, however you may
View all ELISA FAQsReviews for Human HGF DuoSet ELISA
Average Rating: 5 (Based on 2 Reviews)
Have you used Human HGF DuoSet ELISA?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
Filter by: