Mouse IL-2 ELISA Assay Kit

$390.00

The Eagle Biosciences Mouse Interleukin 2 (IL-2) ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of mouse Interleukin 2 (IL-2) concentrations in cell culture supernates, serum, and plasma. The Eagle Biosciences Mouse Interleukin 2 (IL-2) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Mouse IL-2 ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 7 pg/mL
Dynamic Range: 31.25 – 2000 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl

Product manufactured in the USA

Additional Information

Assay Principle

The Eagle Biosciences Mouse Interleukin 2 (IL-2) ELISA Assay Kit employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for interleukin 2 (IL-2) has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any interleukin 2 (IL-2) present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for interleukin 2 (IL-2) is added to the wells and binds to the combination of capture antibody-IL-2 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 interleukin 2 (IL-2) 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 interleukin 2 (IL-2) standard dilutions and interleukin 2 (IL-2) 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 2-8C 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 37C.
  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 37C.
  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 37C 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 37C. 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)

Assay Background

Interleukin 2 (IL-2) is a pleiotropic cytokine produced primarily by mitogen- or antigen-activated T lymphocytes (1, 2). Human IL-2 (also known as T-cell growth factor) is produced by T-cells in response to antigenic or mitogenic stimulation. IL-2 is a potent lymphoid cell growth factor which exerts its biological activity primarily on T cells promoting proliferation and maturation.

Additionally, IL-2 has been found to stimulate growth and differentiation of B cells, NK cells, LAK cells, monocytes, and oligodendocytes. IL-2 is involved in treatment of cancers such as melanoma and renal cell cancer. It plays a key role in promoting the clonal expansion of antigen-specific T cells. In addition, IL-2 has also been shown to mediate multiple immune responses on a variety of cell types.

The sequence of mouse IL-2 cDNA predicts a 169 amino acid (aa) residue   precursor glycoprotein containing a 20 aa residue signal peptide that is cleaved to form the mature protein (3, 4). At the amino acid sequence level, mature mouse IL-2 is approximately 60% identical to human IL-2 (1, 2, 5). Whereas human IL-2 is active on mouse cells, mouse IL-2 is species-specific and is inactive on human cells (6). The gene for IL-2 has been mapped to mouse chromosome 3 (7).  The biological effects of IL-2 are mediated by specific cell surface receptor complexes. The functional high-affinity receptor for IL-2 is composed of three distinct polypeptide chains (8, 9).

IL-2 stimulates the proliferation of thymocytes; stimulates the proliferation and differentiation of activated B cells; promotes the growth, differentiation and cytocidal activity of monocytes; induces the growth of natural killer cells and stimulates cytokine production by these cells as well as the cytolytic activity of these cells; enhances the production of lymphocyte-activated killer (LAK) cells; and induces the proliferation and differentiation of oligodendrocytes (1).

Manual

Product Manual


Publications

References

  • Goldsmith, M.A. and W.C. Greene (1994) in The Cytokine Handbook, 2nd ed., A. Thomson editor, Academic Press, New York, p. 57.
  • Hatakeyama, M. and T. Taniguchi (1990) in Peptide Growth Factors and Their Receptors I, Sporn, M.B. and A.B. Roberts eds., Springer-Verlag, New York, p. 523.
  • Yokota, T. et al. (1985) Proc. Natl. Acd. Sci. USA 82:68.
  • Kashima, N. et al. (1985) Nature 313:401.
  • Taniguchi, T. et al. (1983) Nature 302:305.
  • R&D Systems, Unpublished results.
  • Fuse, A. et al. (1984) Nucl. Acids Res. 12:9323.
  • Kashima, N. et al. (1985) Nature 313:401.
  • Seigel, L.J. et al. (1984) Science 223:175.