Human IL-4 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any 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 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 coloured product is formed in proportion to the amount of 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 IL-2 standard dilutions and IL-2 sample concentration determined.

For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

Interleukin 2 (IL-2) is a pleiotropic cytokine produced primarily by mitogen- or antigen-activated T lymphocytes. 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.

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 the 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 human IL-2 cDNA predicts a 153 amino acid (aa) residue precursor glycoprotein containing a 20 aa residue signal peptide that is cleaved to form the mature protein. At the amino acid sequence level, mouse IL-2 is approximately 60% identical to human IL-2. Whereas human IL-2 is active on mouse cells, mouse IL-2 is species-specific and is inactive on human cells. The gene for IL-2 has been mapped to human chromosome 4q.

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.

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.

GENE ID 3558
SWISS PROT P60568
SYNONYMS Interleukin 2;IL-2;T Cell Growth Factor

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human IL-2 (8﹡12) 1 plate
Human IL-2 Standard lyophilized, 2000 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-human IL-2 monoclonal antibody 2 vials
Streptavidin-HRP solution 2 vials
Standard /sample Diluent 1 bottle
Biotin-Conjugate antibody Diluent 1 bottle
Streptavidin-HRP Diluent 1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20) 1 bottle
Substrate Solution 1 vial
Stop Solution 1 vial
Adhesive Films 4 pieces

Storage

Storage Store at 2 - 8°C

Performance Characteristics

REPEATABILITY The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY The minimum detectable dose was 4pg/mL.
ASSAY RANGE 7.81- 500 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant human IL-2. The factors listed below were prepared at 100ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.
Factors assayed for cross-reactivity
Recombinant human IL-2 sRα, IL-2 Rα, IL-2 Rγ, IL-3, IL-4, IL-6, IL-7, IL-8, LIF, TGF-β1, TGF-β2, TNF-α, TNF-β
Recombinant mouse IL-2, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7
Other proteins bovine FGF acidic, bovine FGF basic, human PDGF, porcine PDGF, human TGF-β1, porcine TGF-β1