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.
Key Product Details
Assay Type
Assay Range
Sample Type
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Mouse 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
Product Summary for Mouse 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 mouse IL-17E. 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 Specifications
Assay Format
Sample Volume Required
Detection Method
Conjugate
Specificity
Label
Scientific Data Images for Mouse IL-17E DuoSet ELISA
Mouse IL-17E / IL-25 ELISA Standard Curve
Kit Contents for Mouse 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
Stability & Storage
Background: IL-17E/IL-25
Long Name
Alternate Names
Gene Symbol
Additional IL-17E/IL-25 Products
Product Documents for Mouse IL-17E DuoSet ELISA
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Mouse IL-17E DuoSet ELISA
For research use only
Related Research Areas
Citations for Mouse IL-17E DuoSet ELISA
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Sample Tested: Bone marrow-derived dendritic cellsVerified Customer | Posted 11/22/2019First time using this kit which has been in the lab for a while. Excellent results. Standards come up very nicely. Instructions clear and easy to follow. 5 stars from me.
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Protocols
View specific protocols for Mouse IL-17E DuoSet ELISA (DY1399):
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.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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- View all Protocols, Troubleshooting, Illustrated assays and Webinars
Associated Pathways
Innate Lymphoid Cell Differentiation Pathways
Th2 Differentiation Pathway
