Human HB-EGF DuoSet ELISA

  (13 citations)     
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Product Details
Assay Procedure
Citations (13)
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    • Assay Type
      Solid Phase Sandwich ELISA
    • Format
      96-well strip plate
    • Sufficient Materials
      For 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.
    Ancillary Reagent Kit Available
    DY008 , DuoSet ELISA Ancillary Reagent Kit 2 - Please Inquire

    This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Heparin Binding EGF-like Growth Factor (HB-EGF). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, saliva, urine, and human milk samples. Diluents for complex matrices, such as serum, plasma, saliva, urine, and human milk, 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)

    Product Datasheets

    Certificate of Analysis Lookup

    To download a Certificate of Analysis, please enter a lot number in the search box below.

    Note: Certificate of Analysis not available for kit components.

    Certificate of Analysis Lookup

    Certificate of Analysis Request Form

    To download a Certificate of Analysis, please enter the catalog and lot numbers in the search box below.

    Note: Certificate of Analysis not available for kit components.

    Preparation and Storage
    • Stability & Storage
      Store the unopened product at 2 - 8 °C. Do not use past expiration date.
    Background: HB-EGF
    HB-EGF (Heparin-Binding EGF-like growth factor), also known as DTR, is produced by bronchial epithelium, visceral and vascular smooth muscle, CD4+ T cells, cardiac muscle, glomerular podocytes, keratinocytes, and IL-10-secreting regulatory macrophages. It activates both the EGF R/ErbB1 and ErbB4 receptors and has been linked to many cellular processes including proliferation, apoptosis, cell migration/invasion, differentiation, morphogenesis, and development. HB-EGF is also involved in several aspects of cancer development and progression.
    • Long Name:
      Heparin Binding EGF-like Growth Factor
    • Entrez Gene IDs:
      1839 (Human); 15200 (Mouse)
    • Alternate Names:
      diphtheria toxin receptor (heparin-binding epidermal growth factor-like growthfactor); Dtr; DTRHEGFLdiphtheria toxin receptor (heparin-binding EGF-like growth factor); Dts; DTSF; HBEGF; HB-EGF; Hegfl; heparin-binding EGF-like growth factor; heparin-binding epidermal growth factor; proheparin-binding EGF-like growth factor
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:

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.

13 Citations: Showing 1 - 10
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Species
Sample Type
  1. BK-UM in patients with recurrent ovarian cancer or peritoneal cancer: a first-in-human phase-I study
    Authors: S Miyamoto, F Yotsumoto, T Ueda, T Fukami, A Sanui, K Miyata, SO Nam, S Fukagawa, T Katsuta, M Maehara, H Kondo, D Miyahara, K Shirota, T Yoshizato, M Kuroki, H Nishikawa, K Saku, Y Tsuboi, K Ishitsuka, Y Takamatsu, K Tamura, A Matsunaga, T Hachisuga, S Nishino, T Odawara, K Maeda, S Manabe, T Ishikawa, Y Okuno, M Ohishi, T Hikita, H Mizushima, R Iwamoto, E Mekada
    BMC Cancer, 2017;17(1):89.
    Species: Human
    Sample Type: Serum
  2. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016;1(13):.
    Species: Human
    Sample Type: Urine
  3. Fingolimod induces neuroprotective factors in human astrocytes.
    Authors: Hoffmann F, Hofereiter J, Rubsamen H, Melms J, Schwarz S, Faber H, Weber P, Putz B, Loleit V, Weber F, Hohlfeld R, Meinl E, Krumbholz M
    J Neuroinflammation, 2015;12(0):184.
    Species: Human
    Sample Type: Cell Culture Supernates
  4. An EGFR wild type-EGFRvIII-HB-EGF feed-forward loop regulates the activation of EGFRvIII.
    Authors: Li L, Chakraborty S, Yang C, Hatanpaa K, Cipher D, Puliyappadamba V, Rehman A, Jiwani A, Mickey B, Madden C, Raisanen J, Burma S, Saha D, Wang Z, Pingle S, Kesari S, Boothman D, Habib A
    Oncogene, 2014;33(33):4253-64.
    Species: Human
    Sample Type: Cell Culture Supernates
  5. Anti-Tumour Effects of a Specific Anti-ADAM17 Antibody in an Ovarian Cancer Model In Vivo.
    Authors: Richards FM, Tape CJ, Jodrell DI, Murphy G
    PLoS ONE, 2012;7(7):e40597.
    Species: Human
    Sample Type: Tissue Homogenates
  6. 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;17(11):3619-30.
    Species: Human
    Sample Type: Complex Sample Type
  7. Assessment of HB-EGF Levels in Peritoneal Fluid and Serum of Ovarian Cancer Patients using ELISA.
    Authors: Hikita S, Yotsumoto F, Fumaki T, Horiuchi S, Sanui A, Miyata K, Nam SO, Tsujioka H, Ueda T, Shirota K, Yoshizato T, Maeda K, Ishikawa T, Okuno Y, Kuroki M, Mekada E, Miyamoto S
    Anticancer Res., 2011;31(7):2553-9.
    Species: Human
    Sample Type: Serum
  8. Heparin binding epidermal growth factor in renal ischaemia/reperfusion injury.
    Authors: Mulder GM, Nijboer WN, Seelen MA, Sandovici M, Bos EM, Melenhorst WB, Trzpis M, Kloosterhuis NJ, Visser L, Henning RH, Leuvenink HG, Ploeg RJ, Sunnarborg SW, van Goor H
    J. Pathol., 2010;221(2):183-92.
    Species: Human
    Sample Type: Urine
  9. Ectodomain shedding of EGFR ligands and TNFR1 dictates hepatocyte apoptosis during fulminant hepatitis in mice.
    Authors: Murthy A, Defamie V, Smookler DS
    J. Clin. Invest., 2010;120(8):2731-44.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  10. IL-17 and IL-22 mediate IL-20 subfamily cytokine production in cultured keratinocytes via increased IL-22 receptor expression.
    Authors: Tohyama M, Hanakawa Y, Shirakata Y, Dai X, Yang L, Hirakawa S, Tokumaru S, Okazaki H, Sayama K, Hashimoto K
    Eur. J. Immunol., 2009;39(10):2779-88.
    Species: Human
    Sample Type: Cell Culture Supernates
  11. Cross-talk between estrogen receptor and epidermal growth factor receptor in head and neck squamous cell carcinoma.
    Authors: Egloff AM, Rothstein ME, Seethala R, Siegfried JM, Grandis JR, Stabile LP
    Clin. Cancer Res., 2009;15(21):6529-40.
    Species: Human
    Sample Type: Cell Culture Supernates
  12. Mining the acute respiratory distress syndrome proteome: identification of the insulin-like growth factor (IGF)/IGF-binding protein-3 pathway in acute lung injury.
    Authors: Schnapp LM, Donohoe S, Chen J, Sunde DA, Kelly PM, Ruzinski J, Martin T, Goodlett DR
    Am. J. Pathol., 2006;169(1):86-95.
    Species: Human
    Sample Type: BALF
  13. 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;10(1):39-50.
    Species: Human
    Sample Type: Cell Culture Supernates
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