Description

Nori Human IL-9 ELISA Kit Summary

Alternative names for IL-9: IL9, interleukin 9,

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 9 (IL-9) is a pleiotropic cytokine (cell signalling molecule) belonging to the group of interleukins.^[1] IL-9 is produced by variety of cells like mast cells, NKT cells, Th2, Th17, Treg, ILC2, and Th9 cells in different amounts. Among them, Th9 cells are regarded as the major CD4+ T cells that produce IL-9. Il-9 is secreted by CD4+ helper cells that acts as a regulator of a variety of hematopoietic cells.^[2] Interleukin 33 (IL-33) induces IL-9 expression and secretion in T cells, which was confirmed by the results obtained in mice by using Human in vitro system.^[3] Whereas the report of others confirms that TGF-β is an essential factor for IL-9 induction.^[4] For the first time (Lars Blom,Britta C. Poulsen,Bettina M. Jensen,Anker Hansen and Lars K. Poulsen published a journal online in 2011 Jul 6),indicating that TGF-β may be important for production of IL-9 but it is not only the definite requirement for IL-9 induction, since cultures with IL-33 without TGF-β have noticeably increased secretion of IL-9, suggesting an important role of IL-33, even though that the effect was not found significant on the gene level.^[5] This cytokine stimulates cell proliferation and prevents apoptosis. It functions through the interleukin-9 receptor (IL9R), which activates different signal transducer and activator (STAT) proteins namely STAT1, STAT3 and STAT5 and thus connects this cytokine to various biological processes. ^[6] The gene encoding this cytokine has been identified as a candidate gene for asthma. Genetic studies on a mouse model of asthma demonstrated that this cytokine is a determining factor in the pathogenesis of bronchial hyperresponsiveness. Interleukin-9 has also shown to inhibit melanoma growth in mice.^[7] Additionally, it gives rise to the multiplication of hematologic neoplasias and also Hodgkin’s lymphoma in humans but IL-9 also has antitumor properties in solid tumors, for example melanoma.

References

1. Kelleher K, et al. (1991). Blood. 77 (7): 1436–41.
2. Perumal NB, Kaplan MH (2011). Trends in Immunology. 32 (4): 146–50.
3. Humphreys NE, et al. (2008). Journal of Immunology. 180 (4): 2443–9.
4. Beriou G, et al. (2010). Journal of Immunology. 185 (1): 46–54.
5. Blom L, et al. (2011). PLoS One. 6 (7): e21695. doi:1371/journal.pone.0021695.
6. Purwar R, et al. (2012). Nature Medicine. 18 (8): 1248–53.
7. Knoops L, Renauld JC (2004). Growth Factors. 22 (4): 207–15.

Product Citation