photo-ELISA Kit
photo-ELISA Kit

Nori Porcine IL-25 ELISA Kit

Nori Porcine IL-11 ELISA Kit
Nori Porcine MMP-8 ELISA Kit
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DataSheet   

This ELISA kit is for quantification of IL-25 in pig. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for IL-25 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-25 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for IL-25 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of IL-25 bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Alternative names for IL-25: Interleukin 25, IL25, interleukin 17E, IL-17E, IL17E

This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.

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Description

Nori Porcine IL-25 ELISA Kit Summary

Alternative names for IL-25: Interleukin 25, IL25, interleukin 17E, IL-17E, IL17E

Alternative names for porcine: pig, swine

Assay Type Solid Phase Sandwich ELISA
Format 96-well Microplate or 96-Well Strip Microplate
Method of Detection Colorimetric
Number of Targets Detected 1
Target Antigen Accession Number A0A8W4FCG8
Assay Length 3 hours
Quantitative/Semiquantitative Quantitative
Sample Type Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL,
Recommended Sample Dilution (Plasma/Serum) No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ
Sensitivity 5 pg/mL
Detection Range 25-1600 pg/mL
Specificity Porcine IL-25
Cross-Reactivity < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested.
Interference No significant interference observed with available related molecules
Storage/Stability 4 ºC for up to 6 months
Usage For Laboratory Research Use Only. Not for diagnostic or therapeutic use.
Additional Notes The kit allows for use in multiple experiments.

 

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Interleukin-25 (IL-25) – also known as IL-17E– is a protein that is encoded by the IL25 gene[1] and contains 177 amino acids. IL-25 is a cytokine that belongs to the IL-17 cytokine family together with IL-17A, IL-17B, IL-17C, IL-17D and IL-17F. IL-25 differs from other members in its function and structure.[2] IL-25 signals through a heterohexameric receptor complex containing IL-17RA and IL-17RB. In this complex, IL-25 forms a homodimer with IL-17RB, which then binds to IL-17RA.[3] The IL-17RA subunit is common for IL-17A and IL-17F, and IL-17RB is common for IL-17B. Both IL-17RA and IL-17RB are essential for IL-25 functions. IL-25 does not bind directly to IL-17RA, but this subunit is necessary for its functions – as well as IL-17RB which directly bind IL-25.[4][5] IL-25 can induce NF-κB activation and production of other cytokines, including IL-4, IL-5, IL-8 and IL-13 in multiple tissues.[1] IL-25 increases IL-9 production in Th9 cells. IL-25 is responsible for the decrease in IFN gamma.[6][7] IL- 25 supports the Th2 immune response and acts to protect against several bowel infections caused by helminths.[7][8]  IL-25 activates natural lymphoid cells 2 (ILC2). IL-25 and IL-33 are the most potent activators of ILC2.[9] IL-25 has been implicated in chronic inflammation associated with the gastrointestinal tract[7]. IL-25 is produced by many cell types and can kill some types of breast cancer cells.[10]

References

  1. Lee J, et al. (2001). The Journal of Biological Chemistry. 276 (2): 1660–4.
  2. Moseley TA, et al. (2003). Cytokine & Growth Factor Reviews. 14 (2): 155–74.
  3. Wilson SC, et al. (2022). Nature. 609 (7927): 622–629.
  4. Rickel EA, et al. (2008). Journal of Immunology. 181 (6): 4299–4310.
  5. Reynolds JM, et al. (2015). Immunity. 42 (4): 692–703.
  6. Angkasekwinai P, et al. (2007). The Journal of Experimental Medicine. 204 (7): 1509–17.
  7. Owyang AM, et al. (2006). The Journal of Experimental Medicine. 203 (4): 843–9.
  8. Pei C, et al. (2016). Infection and Immunity. 84 (12): 3328–3337.
  9. Neill DR, et al. (2010). Nature. 464 (7293): 1367–70.
  10. Furuta S, et al. (2011). Science Translational Medicine. 3 (78): 78ra31.

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