Human EGF 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 EGF. 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: EGF
EGF (Epidermal Growth Factor) is expressed as a transmembrane protein that is proteolytically cleaved to generate soluble forms. It signals through EGF R/ErbB1 which heterodimerizes with ErbB2, ErbB3, or ErbB4 and can also associate with PDGF R beta and HGF R/c-MET. During development, EGF regulates thymocyte differentiation, neuroglia production, and adipocyte maturation. In the adult, EGF plays a role in mammary gland lactogenesis, fibroblast mitosis, dissociation of the extracellular matrix, and cell migration.
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 EGF 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.
21
Citations: Showing 1 - 10
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Lack of Association between Epidermal Growth Factor or Its Receptor and Reflux Esophagitis, Barrett's Esophagus, and Esophageal Adenocarcinoma: A Case-Control Study
Authors: T Deissova, M Cvanova, Z Kala, Z Jiraskova, J Dolina, L Kunovsky, R Kroupa, Z Pavlovsky, B Lipovy, Z Danek, L Izakovicov, O Urban, V Navratil, R Lischke, T Harustiak, T Grolich, V Prochazka, O Slaby, P Borilova L
Disease Markers, 2022-08-31;2022(0):8790748.
Species: Human
Sample Types: Plasma
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Growth Factor Release within Liquid and Solid PRF
Authors: K Zwittnig, B Kirnbauer, N Jakse, P Schlenke, I Mischak, S Ghanaati, S Al-Maawi, D Végh, M Payer, TA Zrnc
Journal of Clinical Medicine, 2022-08-29;11(17):.
Species: Human
Sample Types: Serum
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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
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Effects of the media conditioned by various macrophage subtypes derived from THP-1 cells on tunneling nanotube formation in pancreatic cancer cells
Authors: CW Lee, CC Kuo, CJ Liang, HJ Pan, CN Shen, CH Lee
Oncogene, 2022-07-06;23(1):26.
Species: Human
Sample Types: Cell Culture Supernates
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Preparation and growth factor characterization of cord blood-derived plasma, serum, growth factor-rich plasma and induced serum
Authors: ME Rhéaume, J Perreault, D Fournier, P Trépanier
Cytokine, 2021-11-08;149(0):155756.
Species: Human
Sample Types: Serum
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Frequency Shift of a SH-SAW Biosensor with Glutaraldehyde and 3-Aminopropyltriethoxysilane Functionalized Films for Detection of Epidermal Growth Factor
Authors: XC Lo, JY Li, MT Lee, DJ Yao
Biosensors (Basel), 2020-08-05;10(8):.
Species: Human
Sample Types: Protein
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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
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The neuroprotective activity of heat-treated human platelet lysate biomaterials manufactured from outdated pathogen-reduced (amotosalen/UVA) platelet concentrates
Authors: O Nebie, D Devos, V Vingtdeux, L Barro, JC Devedjian, A Jonneaux, ML Chou, R Bordet, L Buée, F Knutson, D Blum, T Burnouf
J. Biomed. Sci., 2019-10-31;26(1):89.
Species: Human
Sample Types: Cell Culture Supernates
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HER-family ligands promote acquired resistance to trastuzumab in gastric cancer
Authors: A Sampera, FJ Sánchez-Ma, O Arpí, L Visa, M Iglesias, S Menéndez, É Gaye, A Dalmases, S Clavé, M Gelabert-B, TT Poulsen, M Kragh, B Bellosillo, J Albanell, A Rovira, C Montagut
Mol. Cancer Ther., 2019-09-04;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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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
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Tumor associated macrophages induce epithelial to mesenchymal transition via the EGFR/ERK1/2 pathway in head and neck squamous cell carcinoma
Authors: L Gao, W Zhang, WQ Zhong, ZJ Liu, HM Li, ZL Yu, YF Zhao
Oncol. Rep., 2018-08-17;40(5):2558-2572.
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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
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Five-weekly S-1 plus cisplatin therapy combined with trastuzumab therapy in HER2-positive gastric cancer: a phase II trial and biomarker study (WJOG7212G)
Authors: Y Miura, Y Sukawa, S Hironaka, M Mori, K Nishikawa, S Tokunaga, H Okuda, T Sakamoto, K Taku, K Nishikawa, T Moriwaki, Y Negoro, Y Kimura, K Uchino, K Shinozaki, H Shinozaki, N Musha, H Yoshiyama, T Tsuda, Y Miyata, N Sugimoto, T Shirakawa, M Ito, K Yonesaka, K Yoshimura, N Boku, K Nosho, T Takano, I Hyodo
Gastric Cancer, 2017-05-11;0(0):.
Species: Human
Sample Types: Serum
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Reduction of relative centrifugal forces increases growth factor release within solid platelet-rich-fibrin (PRF)-based matrices: a proof of concept of LSCC (low speed centrifugation concept)
Authors: K El Bagdadi, A Kubesch, X Yu, S Al-Maawi, A Orlowska, A Dias, P Booms, E Dohle, R Sader, CJ Kirkpatric, J Choukroun, S Ghanaati
Eur J Trauma Emerg Surg, 2017-03-21;0(0):.
Species: Human
Sample Types: Complex Sample Type
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The EGFR ligands amphiregulin and heparin-binding egf-like growth factor promote peritoneal carcinomatosis in CXCR4-expressing gastric cancer.
Authors: Yasumoto K, Yamada T, Kawashima A, Wang W, Li Q, Donev IS, Tacheuchi S, Mouri H, Yamashita K, Ohtsubo K, Yano S
Clin. Cancer Res., 2011-04-11;17(11):3619-30.
Species: Human
Sample Types: Complex Sample Type
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Identification of biomarkers in human head and neck tumor cell lines that predict for in vitro sensitivity to gefitinib.
Authors: Hickinson DM, Marshall GB, Beran GJ
Clin Transl Sci, 2009-06-01;2(3):183-92.
Species: Human
Sample Types: Cell Lysates
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Development and validation of sandwich ELISA microarrays with minimal assay interference.
Authors: Gonzalez RM, Seurynck-Servoss SL, Crowley SA
J. Proteome Res., 2008-04-19;7(6):2406-14.
Species: Human
Sample Types: Serum
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Amphiregulin is much more abundantly expressed than transforming growth factor-alpha and epidermal growth factor in human follicular fluid obtained from patients undergoing in vitro fertilization-embryo transfer.
Authors: Inoue Y, Miyamoto S, Fukami T, Shirota K, Yotsumoto F, Kawarabayashi T
Fertil. Steril., 2008-03-05;91(4):1035-41.
Species: Human
Sample Types: Follicular Fluid
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Validation of HB-EGF and amphiregulin as targets for human cancer therapy.
Authors: Yotsumoto F, Yagi H, Suzuki SO, Oki E, Tsujioka H, Hachisuga T, Sonoda K, Kawarabayashi T, Mekada E, Miyamoto S
Biochem. Biophys. Res. Commun., 2007-11-20;365(3):555-61.
Species: Human
Sample Types: Cell Culture Supernates
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In-frame deletion in the EGF receptor alters kinase inhibition by gefitinib.
Authors: Sakai K, Yokote H, Murakami-Murofushi K, Tamura T, Saijo N, Nishio K
Biochem. J., 2006-08-01;397(3):537-43.
Species: Human
Sample Types: Cell Culture Supernates
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Targeting ADAM-mediated ligand cleavage to inhibit HER3 and EGFR pathways in non-small cell lung cancer.
Authors: Zhou BB, Peyton M, He B, Liu C, Girard L, Caudler E, Lo Y, Baribaud F, Mikami I, Reguart N, Yang G, Li Y, Yao W, Vaddi K, Gazdar AF, Friedman SM, Jablons DM, Newton RC, Fridman JS, Minna JD, Scherle PA
Cancer Cell, 2006-07-01;10(1):39-50.
Species: Human
Sample Types: Cell Culture Supernates
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