Human IL-22 ELISA Assay

$445.00

The Human IL-22 ELISA Assay (enzyme-linked immunoassay kit) is intended for the quantitative determination of human Interleukin 22 (IL-22) concentrations in cell culture supernates, serum, and plasma. The Eagle Biosciences Human Interleukin 22 (IL-22) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: L2231-K01 Categories: , ,

Human IL-22 ELISA Assay

The Human IL-22 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 15 pg/mL
Dynamic Range: 31.25 – 1000 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: Interleukin 22, IL-10-related T cell-derived inducible factor, IL-TIF, IL10 TIF


Assay Background

Interleukin 22 (IL-22), also known as IL-10-related T cell-derived inducible factor (IL-TIF), is a member of the IL-10 cytokine family. Other members of this family include IL-10, IL-19, IL-20, IL-24, and IL-26 (1). IL-22 was initially identified as a gene induced by IL-9 in mouse T cells and mast cells. Human IL-22 cDNA encodes a 179 amino acid (aa) protein with a putative
33 aa signal peptide, sharing approximately 79% and 22% aa sequence identity with mouse IL-22 and human IL-10, respectively. Although the related IL-10 is thought to act as a dimer, the crystal structure of IL-22 suggests it may interact with its receptor as a monomer.  The functional IL-22 receptor is of the class 2 subtype and consists of two receptor subunits,IL-22 R (previously an orphan receptor named CRF2-9) and IL-10 R  (previously known as CRF2-4). The IL-10 R chain is shared by IL-10, IL-26, IL-28A, IL-28B, and IL-29.
IL-22 R is expressed primarily in the pancreas, and to a lesser extent, tissues of the gastrointestinal tract, kidney, and skin. A soluble receptor, IL-22 binding protein (IL-22BP), has also been described and may act as an endogenous inhibitor of IL-22 activity. IL-22 has been shown to activate Jak/STAT and MAPK signaling pathways and upregulate the production of acute phase proteins.

IL-22 is produced primarily by activated Th1-type T cells and NK cells (19). Mouse IL-22 expression is induced in various organs upon lipopolysaccharide injection, suggesting that it may be involved in inflammatory responses (3). In humans, this is supported by the observation that IL-22 is produced by synovial fibroblasts and macrophages of rheumatoid arthritis (RA) patients and is capable of inducing pro-inflammatory responses in RA synovial tissues. In addition, it stimulates the production of pro-inflammatory cytokines and anti-microbial defensins in human keratinocytes. These activities result in epidermal hyperplasia in models of human skin.


Related Products

Human IL-21 ELISA Assay
Human IL-32 Alpha ELISA Assay Kit

Additional Information

Assay Principle


The Eagle Biosciences Human Interleukin 22 (IL-22) ELISA Assay Kit employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-22 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-22 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-22 is added to the wells and binds to the combination of capture antibody- IL-22 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A colored product is formed in proportion to the amount of IL-22 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-22 standard dilutions and IL-22 sample concentration determined.

  1. Prepare all reagents and working standards as directed in the previous sections.
  2. Determine the number of microwell strips required to test the desired number of samples plus appropriate number of wells needed for running blanks and standards. Remove extra microwell strips from holder and store in foil bag with the desiccant provided at 2-8°C sealed tightly.
  3. Add 100 µL of Standard, control, or sample, per well. Cover with the adhesive strip provided. Incubate for 1.5 hours at 37°C.
  4. Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with Wash Buffer (350 µL) using a squirt bottle, manifold dispenser or auto-washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
  5. Add 100 µL of the working solution of Biotin-Conjugate to each well. Cover with a new adhesive strip and incubate 1 hour at 37°C.
  6. Repeat the aspiration/wash.
  7. Add 100 µL of the working solution of Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at 37°C Avoid placing the plate in direct light.
  8. Repeat the aspiration/wash.
  9. Add 100 µL of Substrate Solution to each well. Incubate for 10-20 minutes at 37°C. Avoid placing the plate in direct light.
  10. Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  11. Determine the optical density of each well immediately, using a microplate reader set to 450 nm.(optionally 630nm as the reference wave length;610-650nm is acceptable) 

Documents

Product Manual


 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

Publications

References


1. Pestka, S. et al. (2004) Annu. Rev. Immunol. 22:929.
2. Dumoutier, L. et al. (2000) J. Immunol. 164:1814.
3. Dumoutier, L. et al. (2000) Proc. Natl. Acad. Sci. USA 97:10144.
4. Xie, M.H. et al. (2000) J. Biol. Chem. 275:31335.
5. Nagem, R.A. et al. (2002) Acta Crystallogr. D. Biol. Crystallogr. 58:529.
6. Kotenko, S.V. et al. (2001) J. Biol. Chem. 276:2725.
7. Kotenko, S.V. and J.A. Langer. (2004) Int. Immunopharmacol. 4:593.
8. Donnelly, R.P. et al. (2004) J. Leukoc. Biol. 76:314.
9. Boniface, K. et al. (2005) J. Immunol. 174:3695.
10. Wolk, K. et al. (2004) Immunity 21:241.
11. Gurney, A.L. (2004) Int. Immunopharmacol. 4:669.
12. Aggarwal, S. et al. (2001) J. Interferon Cytokine Res. 21:1047.
13. Dumoutier, L. et al. (2001) J. Immunol. 166:7090.
14. Kotenko, S.V. et al. (2001) J. Immunol. 166:7096.
15. Xu, W. et al. (2001) Proc. Natl. Acad. Sci. USA 98:9511.
16. Nagalakshmi, M.L. et al. (2004) Int. Immunopharmacol. 4:679.
17. Lejeune, D. et al. (2002) J. Biol. Chem. 277:33676.
18. Dumoutier, L. et al. (2000) Genes Immun. 1:488.
19. Wolk, K. et al. (2002) J. Immunol. 168:5397.
20. Ikeuchi, H. et al. (2005) Arthritis Rheum. 52:1037.9.

Product Citations