There are five classes of mammalian immunoglobulins: IgA, IgD, IgE, IgG, and IgM. In mice, the IgG class is further divided into four subclasses: IgG1, IgG2a/ IgG2c (strain specific), IgG2b, and IgG3.The general immunoglobulin structure is described as two heavy and two light chain (H2L2) polypeptides linked together by disulfide bridges of cysteine residues. Mouse IgE, also known as reaginic antibody, exists in serum as a 4-chain monomer of 185–200 kDa. Like IgM, the IgE monomers contain a fourth constant region domain. Serum concentrations of IgE are reported to be >10 ug/ml. IgE is cyolytic for mast cells, where it binds to the FcRI receptor and is able to mediate the response of granular and lipid mediators of inflammation, including the PCA reaction. The IgE class switch is induced by IL-4 as part of the Th2 response.

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IgE has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps，any IgE present is bound by the immobilized antibody and the detection antibody specific for IgE is binds to the combination of capture antibody- IgE 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 IgE 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 IgE standard dilutions and IgE sample concentration determined.

Figure 1:Schematic diagram of the assay

1. Aluminium pouches with a Microwell Plate coated with antibody to mouse IgE (812)

2. 2 vials mouse IgE Standard lyophilized, 40000 pg/ml upon reconstitution

3. 2 vials concentrated Biotin-Conjugate anti-mouse IgE antibody

4. 2 vials Streptavidin-HRP solution

5. 1 bottle Standard /sample Diluent

6. 1 bottle Biotin-Conjugate antibody Diluent

7. 1 bottle Streptavidin-HRP Diluent

8. 1 bottle Wash Buffer Concentrate 20x (PBS with 1% Tween-20)

9. 1 vial Substrate Solution

10. 1 vial Stop Solution

11. 4 pieces Adhesive Films

12. package insert

NOTE: [96 Tests]

Unopened Kit：Store at 2 -8° C. Do not use past kit expiration date. opened/ReconstitutedReagents：Please refer to the datasheets for detail information.

1. Quimby, F. W. & Luong Richard, H. (2007). Clinical Chemistry of the Laboratory Mouse. In

J.G.Fox, S. W. Barthold, M. T. Davisson, C. E. Newcomer, F. W. Quimby, & A. L. Smith (Eds.), The Mouse in Biomedical Research: Vol. III Normative Biology, Husbandry, and Models (pp. 190). Burlington: Academic Press.