Human IL-17E DuoSet ELISA

R&D Systems | Catalog # DY1258-05

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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

11.7-750 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human IL-17E DuoSet ELISA 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
View all IL-17E/IL-25 ELISA Kits »

Product Summary for Human IL-17E DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Interleukin 17E (IL-17E). 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 Specifications

Assay Format

96-well strip plate (sold separately)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human IL-17E DuoSet ELISA

Human IL-17E / IL-25 ELISA Standard Curve

Human IL-17E / IL-25 ELISA Standard Curve

Kit Contents for Human IL-17E DuoSet ELISA

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

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 equivalent

Reagent Diluent*

Blocking Buffer*

Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)

Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)

Microplates: (Catalog # DY990), or equivalent

Plate Sealers: (Catalog # DY992), or equivalent

*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.

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: IL-17E/IL-25

IL-17E/IL-25 (Interleukin-25) is a cytokine that promotes Th2-biased immune responses. This is in contrast to other IL-17 family members which promote Th1- and Th17-biased inflammation. IL-25 is an important mediator of allergic reactions and protection against intestinal parasites. During helminth infections and allergic reactions, IL-25 is locally upregulated in intestinal and airway epithelial cells, atopic dermatitis skin lesions, and local Th2 cells, eosinophils, and basophils. It binds to IL-17 RB but also requires IL-17 RA to exert its activity. IL-25 acts on a variety of cell types which respond with increased production of Th2 cytokines (e.g. IL-4, IL-5, IL-13) and reduced production of Th1 and Th17 cytokines (e.g. IFN-gamma, IL-12, IL-23, IL-17A, IL-17F). Airway IL-25 can be activated by MMP-7, a protease that is upregulated in airway epithelium in response to allergen exposure. Cleaved IL-25 shows enhanced binding to IL-17 RB and stronger induction of Th2 cytokines. IL-25 also promotes vascular endothelial cell proliferation and assembly into tubular structures. It supports the integrity of the blood-brain barrier and limits CD4+ T cell infiltration into the brain.

Long Name

Interleukin 17E

Alternate Names

IL-25, IL17E, IL25

Entrez Gene IDs

64806 (Human); 140806 (Mouse); 501996 (Rat)

Gene Symbol

IL25

Additional IL-17E/IL-25 Products

Product Documents for Human IL-17E DuoSet ELISA

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human IL-17E DuoSet ELISA

For research use only

Citations for Human IL-17E DuoSet ELISA

Customer Reviews for Human IL-17E DuoSet ELISA (3)

4.3 out of 5
3 Customer Ratings
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  • Human IL-17E DuoSet ELISA
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 04/02/2020
    We used this kit for the quantification of IL-17e in human serum and plasma. Works very well and well-described protocol.
    Human IL-17E DuoSet ELISA DY1258-05
  • Human IL-17E DuoSet ELISA
    Name: Anonymous
    Sample Tested: Human serum
    Verified Customer | Posted 05/08/2017
    Human IL-17E DuoSet ELISA DY1258-05
  • Human IL-17E DuoSet ELISA
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 05/01/2017
    Good
    Human IL-17E DuoSet ELISA DY1258-05

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Protocols

View specific protocols for Human IL-17E DuoSet ELISA (DY1258-05):

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.

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