Overview

Introduction
Eotaxin, also known as Chemokine (C-C motif) ligand 11 (CCL11), is a small cytokine belonging to the CC chemokine family. These family members play an important role in the recruitment of leukocytes to inflammatory lesions. Eotaxin is a potent inducer of eosinophil chemotaxis leading to eosinophil migration in vitro and accumulation in vivo. It also interacts with CCR2 and CCR5 and can thus affect the responses of monocytes, which express both receptors. CCL11 selectively recruits eosinophils by inducing their chemotaxis, and therefore, is implicated in allergic responses. It has been found that eotaxin has high degree selectivity for its receptor, such that they are inactive on neutrophils and monocytes, which do not express CCR3.

Principle of the Assay  
This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Eotaxin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Eotaxin polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were 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 Eotaxin amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Eotaxin can be calculated.

Range: 31.2 pg/ml-2000 pg/ml

Sensitivity < 2 pg/ml

Storage and Expiration: Store at 2-8℃ for 4 months, or at -20℃ for 8 months.

Application: For quantitative detection of Eotaxin in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Kit components
1.     One 96-well plate pre-coated with anti-Human Eotaxin antibody
2.     Lyophilized Human Eotaxin standards: 2 tubes (10ng / tube)                    
3.     Sample / Standard diluent buffer: 30 ml
4.     Biotin conjugated anti-Human Eotaxin antibody (Concentrated): 130 μl. Dilution: 1:100
5.     Antibody diluent buffer: 12 ml
6.     Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130 μl. Dilution: 1:100
7.     ABC diluent buffer: 12 ml
8.     TMB substrate: 10 ml
9.     Stop solution: 10 ml
10.   Wash buffer (25X): 30 ml

Note: Reconstitute standards and test samples with Kit Component 3. 

Gene Information

OMIM: 601156

Swiss Port: P51671
Gene ID: 6356

Other Information

Please check datasheet for protocol.