Mouse IL-5 is a heamotapoietic cytokine, mainly produced by T-helper 2 cells and mast cells. The cytokine is comprised of two identical 113 amino acid units linked by disulfide bond. The gene for mouse IL-5 is located at chromosome 11 in conjunction with genes for IL4 and other T-helper 2 cytokines such as IL-13, GM-CSF, IL-3. IL-5 regulates B cell proliferation, eosinophil maturation and activation. IL-5 receptor is a heterodimer with a cytokine specific alpha unit, and a signal transduction beta subunit that is identical to the beta subunit of receptors for IL-3 and GM-CSF. IL-5 receptor alpha (IL-5Ra) is expressed by eosinophils, B cells and basophils in member-bound form and soluble form. While the membrane bound form participates in the activation of eosinophils and B cells, the soluble form of IL-5Ra has an antagonist effect to IL-5. Increased IL-5 expression causes chronic IgE response and hypereosinophilic syndromes. Elevated IL-5 level is found in allergic diseases such as allergic rhinitis, asthma and allergic conjunctivitis. This ELISA kit provides a tool for studying IL-5 expression and regulation in animal model.
Intended Use:This Mouse IL-5 ELISA kit is to be used for the in vitro quantitative determination of mouse interleukin 5 (IL-5) concentrations in serum, cell culture supernatant, and other biological fluids.
Sensitivity:<0.8pg/ml
Range:3.125-200pg/ml
Test Principle:This mouse IL-5 enzyme-linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific for mouse IL-5. When standards or samples are added to the appropriate microtiter plate wells, mouse IL-5 in the standards or samples will be immobilized by the precoated antibody during incubation. Then, a biotin-conjugated antibody preparation specific for mouse IL-5 is added to each well and incubated. The biotin labelled antibody attaches to the wells by binding to mouse IL-5. After plate washing, other proteins, components and unattached biotin labelled antibody is removed. After that, avidin-horseradish peroxidase (HRP) conjugate is added to each well. Avidin has a very high affinity for biotin, thus, it links the tracer (HRP) sturdily to the biotin labelled antibody. The wells are thoroughly washed to remove all unbound avidin-HRPconjugate and aTMB(3,3’, 5,5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only wells that contain mouse IL-5 will exhibit a change in color. The extent of color change is proportional to the quantity of mouse IL-5 present in the standards/samples. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wave length of 450nm ± 2nm
In order to measure the concentration of mouse IL-5 in the samples, this kit contains two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing). According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D.) versus IL-5 concentration (pg/ml). The concentration of mouse IL-5 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Kit Components:MOUSE IL-5 MICROTITER PLATE BIOTIN CONJUGATE AVIDIN CONJUGATE MOUSE IL-5 STANDARD CALIBRATOR DILUENT I CALIBRATOR DILUENT II WASH BUFFER (20X/96 wells, 30X/192 wells) TMB SUBSTRATE STOP SOLUTION
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
