Mouse HB-EGF DuoSet ELISA

Catalog # Availability Size / Price Qty
DY8239-05
Ancillary Products Available
Mouse HB-EGF ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (1)
FAQs
Supplemental Products
Reviews (2)

Mouse HB-EGF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For five 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 mouse Heparin Binding EGF-like Growth Factor (HB-EGF). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma 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)

Data Example

Mouse HB-EGF ELISA Standard Curve

Product Datasheets

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: 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 of Block Buffer 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.

Citation for Mouse HB-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.

1 Citation: Showing 1 - 1

  1. Deoxycholic acid activates epidermal growth factor receptor and promotes intestinal carcinogenesis by ADAM17-dependent ligand release
    Authors: W Dong, L Liu, Y Dou, M Xu, T Liu, S Wang, Y Zhang, B Deng, B Wang, H Cao
    J. Cell. Mol. Med., 2018;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates

FAQs

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Reviews for Mouse HB-EGF DuoSet ELISA

Average Rating: 2.5 (Based on 2 Reviews)

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Mouse HB-EGF DuoSet ELISA
By Anonymous on 04/29/2021
Application: Sample Tested: Bronchoalveolar lavage,Lung homogenate

Bottom half of standard curve completely unusable, didn't develop past 4th point on standard curve. This happened across several plates and on three separate occasions. Top OD reached after ~20 mins of incubation with TMB was only about 0.5 (normally 1-2.5).

HB-EGF ELISA was done in parallel to mouse AREG, BTC and EREG and those all worked fine. All reagents were made up in parallel as well.


Mouse HB-EGF DuoSet ELISA
By Anonymous on 07/19/2018
Application: Sample Tested: eye tissue

protocol was fairly easy to follow (you have to keep going back and forth between the handout and the CoA sheet provided in order to make the reagents properly). Unfortunately, there was not a positive control, so I had to use recombinant protein to spike the assay to make sure it worked properly.