Nori Mouse IL-22 ELISA Kit Summary

Alternative names for IL-22: Interleukin 22

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-22 (IL-22) is protein that is encoded by the IL22 gene.^[1] IL-22 is an α-helical cytokine. IL-22 binds to a heterodimeric cell surface receptor composed of IL-10R2 and IL-22R1subunits.^[2] IL-22 is a member of the IL-10 family,^[3] a class of potent mediators of cellular inflammatory responses. It shares use of IL-10R2 in cell signaling with other members of this family, IL-10, IL-26, IL-28A/B and IL-29.^[4] IL-22 is produced by activated DC and T cells and initiates innate immune responses against bacterial pathogens especially in epithelial cells such as respiratory and gut epithelial cells. IL-22 along with IL-17 is rapidly produced by splenic LTi-like cells ^[5] and also produced by Th17 cells and likely plays a role in the coordinated response of both adaptive innate immune systems, autoimmunity and tissue regeneration.^[6] IL-22 biological activity is initiated by binding to a cell-surface complex composed of IL-22R1 and IL-10R2 receptor chains and further regulated by interactions with a soluble binding protein, IL-22BP, which shares sequence similarity with an extracellular region of IL-22R1 (sIL-22R1). IL-22 and IL-10 receptor chains play a role in cellular targeting and signal transduction to selectively initiate and regulate immune responses.^[2] IL-22 can contribute to immune disease through the stimulation of inflammatory responses, S100s and defensins. IL-22 also promotes hepatocyte survival in the liver and epithelial cells in the lung and gut similar to IL-10.^[7] In some contexts, the pro-inflammatory versus tissue-protective functions of IL-22 are regulated by the often co-expressed cytokine IL-17A ^[8]

References

1. Xie MH, et al. (2000). The Journal of Biological Chemistry. 275(40): 31335–9.
2. Jones BC, et al. (2008). Structure. 16(9): 1333–44.
3. Pestka S, et al. (2004). Annual Review of Immunology. 22: 929–79.
4. Witte K, et al. (2010). Cytokine & Growth Factor Reviews. 21(4): 237–51.
5. Takatori H, et al. (2009).  The Journal of Experimental Medicine. 206(1): 35–41.
6. Nikoopour E, et al. (2015). Cytokine. 74(1): 35–42. doi:1016/j.cyto.2014.09.007.
7. Moore KW, et al. (2001). Annual Review of Immunology. 19: 683–765.
8. Sonnenberg GF, et al. (2010).  The Journal of Experimental Medicine. 207(6): 1293–305.