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Human EGF DuoSet ELISA

Catalog #: DY236
21 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY236
DY236-05
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human EGF ELISA Standard Curve
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Product Details
Procedure
Citations (21)
FAQs
Supplemental Products
Reviews (1)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human EGF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
3.9 - 250 pg/mL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* 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

N/A Human EGF ELISA Standard Curve View Larger

Human EGF ELISA Standard Curve

Product Datasheets

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Product Datasheet
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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: 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.

Long Name:
Epidermal Growth Factor
Entrez Gene IDs:
1950 (Human); 13645 (Mouse); 25313 (Rat)
Alternate Names:
beta-urogastrone; EGF; epidermal growth factor (beta-urogastrone); epidermal growth factor; hEGF; HOMG4; pro-epidermal growth factor; URG; Urogastrone

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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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  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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.
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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
  21. 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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Human EGF DuoSet ELISA
By Anonymous on 11/23/2017
Sample Tested: Various cell lines

Procedures explained with clarity and kit comes with excellent quality materials for excellent data collection. Be thorough and careful with concentration calculations

Human EGF DuoSet ELISA DY236