Description

Nori Sheep IL-10 ELISA Kit Summary

Alternative names for IL-10: IL10, interleukin 10, cytokine synthesis inhibitory factor (CSIF),

Alternative name for sheep: ovine, lamb, goat

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: 

IL-10, also known as cytokine synthesis inhibitory factor (CSIF), is an anti-inflammatory cytokine that is produced by T cells, NK cells, mast cells and macrophages (1,2,3). It is capable of inhibiting synthesis of pro-inflammatory cytokines like IFN-γ, IL-2, IL-3, TNFα and GM-CSF made by cells such as macrophages and regulatory T-cells. IL-10 also displays potent abilities to suppress the antigen presentation capacity of antigen presenting cells. IL-10 initiates signal transduction by binding to a cell surface receptor complex consisting of IL-10 RI and IL-10 RII (1). Binding of IL-10 leads to the activation of Jak1 and Tyk2, which phosphorylates Stat-3 (1,4). The anti-inflammatory activity of IL-10 is due to its ability to block signaling through other cytokine receptors, notably IFNγ receptor, by upregulating expression of SOCS-1 (1,4). In addition, IL-10 promotes T cell tolerance by inhibiting tyrosine phosphorylation of CD28 (5,6). IL-10 is an important negative regulator of the immune response, which allows for maintenance of pregnancy (1). In contrast, increased IL-10 levels contribute to persistent Leishmania major infections (7).

References

1. Pestka, S. et al. (2004) Immunol Rev 202, 8-32.
2. Akuffo, H. et al. (1999) Clin Exp Immunol 117, 529-34.
3. Grimbaldeston MA, et al (2007). Nat. Immunol. 8 (10): 1095–104.
4. O’Shea, J.J. and Murray, P.J. (2008) Immunity 28, 477-87.
5. Akdis, C.A. and Blaser, K. (2001) Immunology 103, 131-6.
6. Akdis, C.A. et al. (2000) FASEB J 14, 1666-8.
7. Von Stebut, E. (2000) Eur J Dermatol 17, 115-22.

Product Citation