Mouse IgE ELISA Assay Kit

$390.00

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

TGF-Beta 1 ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 156 pg/mL
Dynamic Range: 312.5 – 10,000 pg/ml
Incubation Time: 3 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl

Product manufactured in the USA

Additional Information

Assay Principle

The Eagle Biosciences Mouse IgE ELISA Assay Kit 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 colored 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.

  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 2-8°C sealed tightly.
  3. Add 100 µl of Standard, control, or sample, per well, then add 50 µl of the working solution of Biotin-Conjugate to each well. Cover with the adhesive strip provided and incubate 2 hours at RT. Adequate mixing is very important for good result. Use a mini-vortexer at the lowest frequency.
  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 Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at RT Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash.
  7. Add 100 µl of Substrate Solution to each well. Incubate for 10-20 minutes at RT. Avoid placing the plate in direct light.
  8. Add 100 µl of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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

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 FcRI 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.

Manual

Product Manual


Publications

References

1. Sims, J.E. et al. (2000) J. Exp. Med. 192:671.
2. Park, L.S. et al. (2000) J. Exp. Med. 192:659.
3. Pandey, A. et al. (2000) Nature Immunol. 1:59.
4. Reche, P.A. et al. (2001) J. Immunol. 167:336.