Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-4 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-4 is added to the wells and binds to the combination of capture antibody-IL-4 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-4 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-4 standard dilutions and IL-4 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interleukin 4 (IL-4) is a pleiotropic cytokine produced primarily by activated T lymphocytes, mast cells and basophils.
The cDNA sequence of mouse IL-4 predicts a 140 amino acid (aa) residue precursor protein containing a 20 aa residue signal peptide that is cleaved to form the mature protein. At the amino acid sequence level, mature mouse IL-4 is approximately 50% identical to human IL-4 but there is no species cross-reactivity for biological activity for the two proteins. Mouse IL-4 also shares approximately 30% amino acid sequence identity to mouse IL-13 and the two cytokines exhibit overlapping biological activities. The gene for IL-4 has been mapped to mouse chromosome 11, in close proximity to the genes for IL-3, IL-5, IL-13 and GM-CSF.
IL-4 has multiple immune response-modulating activities on a variety of cell types. It is an important regulator of isotype switching, inducing IgE production in B lymphocytes. It is an important modulator of the differentiation of precursor T helper cells to the Th2 subset that mediates humoral immunity and modulates antibody production. In addition, IL-4 has also been shown to have anti-tumor activity both in vivo and in vitro.
The biological effects of IL-4 are mediated by specific cell surface receptor complexes. Although IL-4 R does not bind IL-13 directly, it has been shown to complex with the low-affinity IL-13 R to form the functional high-affinity receptor complex for IL-13. In addition to the membrane-bound form of IL-4 R, a naturally occurring soluble form of IL-4 R has been identified in human and mouse biological fluids and in mouse cell culture supernates. Soluble IL-4 R has been to shown to bind IL-4 with high affinity in solution.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to mouse IL-4(8﹡12)||1 plate|
|Mouse IL-4 Standard lyophilized, 2000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-mouse IL-4 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|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 7pg/mL.|
|ASSAY RANGE||15.6 - 1000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant mouse IL-4. The factors listed below were prepared at 50ng/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 mouse||G-CSF, GM-CSF, IL-1α, IL-1β, IL-2, IL-3, IL-5, IL-6, IL-7, IL-9, IL-10, IL-13, LIF, VEGF, MIP-2, TNF-α, SCF|
|Other proteins||IL-4, IL-4sRα|
Data Analysis Assistance
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