Description

Nori Equine IgE ELISA Kit Summary

Alternative names for IgE: Immunoglobulin E

Alternative names for equine: horse

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background:

Immunoglobulin E (IgE) is a kind of antibody (or immunoglobulin (Ig) “isotype“) that has only been found in mammals. Monomers of IgE consist of two heavy chains (ε chain) and two light chains, with the ε chain containing 4 Ig-like constant domains (Cε1-Cε4). IgE’s main function is immunity to parasites such as helminths^[1] like Schistosoma mansoni, Trichinella spiralis, and Fasciola hepatica.^[2] IgE is utilized during immune defense against certain protozoan parasites such as Plasmodium falciparum. IgE also has an essential role in type I hypersensitivity,^[3] which manifests various allergic diseases, such as allergic asthma, most types of sinusitis, allergic rhinitis, food allergies, and specific types of chronic urticaria and atopic dermatitis. IgE also plays a pivotal role in responses to allergens, such as: anaphylactic drugs, bee stings, and antigen preparations used in desensitization immunotherapy. Although IgE is typically the least abundant isotype—blood serum IgE levels in a normal (“non-atopic“) individual are only 0.05% of the Ig concentration,^[4] compared to 75% for the IgGs at 10 mg/ml, which are the isotypes responsible for most of the classical adaptive immune response—it is capable of triggering the most powerful inflammatory reactions. IgE primes the IgE-mediated allergic response by binding to Fc receptors found on the surface of mast cells and basophils. IgE may play an important role in the immune system’s recognition of cancer,^[5] in which the stimulation of a strong cytotoxic response against cells displaying only small amounts of early cancer markers would be beneficial. IgE is known to be elevated in various autoimmune disorders such as Lupus(SLE), Rheumatoid Arthritis(RA) & psoriasis, and is theorized to be of pathogenetic importance in RA and SLE by eliciting a hypersensitivity reaction.^[6]

References

1. Erb KJ (2007). Eur. J. Immunol. 37 (5): 1170–3.
2. Pfister K, et al. (1983). “IgE production in rat fascioliasis”. Parasite Immunol. 5 (6): 587–93.
3. Gould HJ, et al. (2003). Annu. Rev. Immunol. 21: 579–628.
4. Winter WE, et al. (2000). Arch. Pathol. Lab. Med. 124 (9): 1382–5.
5. Karagiannis SN, et al. (2003). Eur. J. Immunol. 33 (4): 1030–40.
6. Elkayam O, et al. (1995). Allergy 50 (1): 94–6.

Product Citations