Description
Principle of the assay: human Eotaxin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Eotaxin has been precoated onto 96-well plates. Standards(E.coli,G24-P97) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Eotaxin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Eotaxin amount of sample captured in plate.
Background: Eotaxin, also known as CCL11, is a potent inducer of eosinophil chemotaxis and is considered as a selective ligand of the CC chemokine receptor 3 (CCR3), which is expressed on eosinophils, basophils, and Th2 lymphocytes.1 Eotaxin is assumed to be involved in eosinophilic inflammatory diseases such as atopic dermatitis,allergic rhinitis, asthma and parasitic infections.2 The gene maps to chromosome 17 and is expressedconstitutively at high levels in small intestine and colon, and at lower levels in various other tissues. The deduced mature protein sequence is 66% identical to human monocyte chemoattractant protein-1, and 60% identical to guinea pig eotaxin. Recombinant human eotaxin produced in insect cells induces a calcium flux response in normal human eosinophils, but not in neutrophils or monocytes.3 The human eotaxin gene is cloned and found tobe 61.8 and 63.2% identical at the amino acid level to guinea pig and mouse eotaxin.4