Description

Nori Hamster IL-2 ELISA Kit Summary

Alternative names for IL-2: IL2, interleukin 2, TCGF

Alternative name for hamster: golden hamster, syrian hamster

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-2 is an interleukin, a type of cytokine immune system signaling molecule. IL-2 is a T cell stimulatory cytokine best known for inducing T cell proliferation, the production of immunoglobulins made by B cells and differentiation and proliferation of natural killer cells (1,2). IL-2 is also necessary during T cell development in the thymus for the maturation of a unique subset of T cells that are termed regulatory T cells (T-regs) (3, 4, 5). Conversely, IL-2 is involved in activation induced cell death (AICD) that is observed post T cell expansion by increasing levels of Fas on CD4+ T cells (6). IL-2 signals through a trimeric receptor complex consisting of IL-2 receptor alpha (CD25), IL-2 receptor beta (CD122) and a common gamma chain (γc). IL-2 Rα binds exclusively to IL-2 with low affinity and increases binding affinity of the whole receptor complex including IL-2Rβ and γc subunits. IL-2Rβ is also used by IL-2. The common γc is shared by all members of this family of cytokines including IL-4, IL-7, IL-9, IL-2, and IL-21. Binding of IL-2 activates the Ras/MAPK, JAK/Stat and PI 3-kinase/Akt signaling modules (1,2).

References

1. Ma, A. et al. (2006) Annu Rev Immunol 24, 657-79.
2. Gaffen, S.L. and Liu, K.D. (2004) Cytokine 28, 109-23.
3. Sakaguchi S, et al (1995). J. Immunol. 155 (3): 1151–64.
4. Fehérvari, Z. et al. (2006) Trends Immunol 27, 109-11.
5. Antony, P.A. et al. (2006) J Immunol 176, 5255-66.
6. Jaleco, S. et al. (2003) J Immunol 171, 61-8.

Product Citation