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Human MIP-1a ELISAMacrophage Inflammatory Proteins (MIP) is a member of the C-C subfamily of chemokines that exhibit a variety of proinflammatory activities in vitro including leukocyte chemotaxis. There are two major forms, MIP-1a and MIP-1b, in humans. They are major factors produced by macrophages after stimulated with bacterial endotoxins. They activate human granulocytes (neutrophils, eosinophils and basophils) which can lead to acute neutrophilic inflammation. They also induce the synthesis and release of other pro-inflammatory cytokines such as IL-1, IL-6 and TNFa from fibroblasts and macrophages. In addition to its proinflammatory activities, MIP-1a inhibits the proliferation of hematopoietic stem cells in vitro and in vivo. A. Benefits:
B. Principle of the technology MIP-1a ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay utilizes goat anti-human MIP-1a antibodies for immobilization on the microtiter wells and goat anti-human MIP-1a antibodies along with streptavidin conjugated to horseradish peroxidase (HRP) for detection. The test sample is allowed to react simultaneously with the two antibodies, resulting in the MIP-1a molecules being sandwiched between the solid phase and enzyme-linked antibodies. After incubation, the wells are washed to remove unbound-labeled antibodies. A HRP substrate, TMB, is added to result in the development of a blue color. The color development is then stopped with the addition of Stop Solution changing the color to yellow. The concentration of MIP-1a is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm. Figure 1 Figure 2
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