Description
Introduction: Interferon-inducible protein-10 (IP-10) is a member of the C-X-C chemokine family that is also known as 10 kDa interferon-gamma-induced protein (gamma-IP10 or IP-10). IP-10 is secreted by several cell types in response to IFN-gamma. These cell types include monocytes, endothelial cells and fibroblasts. IP-10 has been attributed to several roles, such as chemoattraction for monocytes/macrophages, T cells, NK cells, and dendritic cells, promotion of T cell adhesion to endothelial cells, antitumor activity, and inhibition of bone marrow colony formation and angiogenesis. The gene for IP-10 is located on human chromosome 4 in a cluster among several other CXC chemokines. This chemokine elicits its effects by binding to the cell surface chemokine receptor CXCR3. The three-dimensional crystal structure of this chemokine has been determined under 3 different conditions to a resolution of up to 1.92A. The Protein Data Bank accession codes for the structures of CXCL10 are 1lv9, 1o7y, 1o7z and 1o80.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to IP-10. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for IP-10 and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain IP-10, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of IP-10 in the samples is then determined by comparing the O.D. of the samples to the standard curve