Description:
Macrophage Inflammatory Protein-1 (MIP-1) is a member of the C-C subfamily of chemokines that exhibit a variety of proinflammatory activities in vitro including leukocyte chemotaxis. MIP-1 is a major factor produced by macrophages after stimulated with bacterial endotoxins. It activates granulocytes (neutrophils, eosinophils and basophils) which can lead to acute neutrophilic inflammation. It also induces 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-1 inhibits the proliferation of hematopoietic stem cells in vitro and in vivo.Principle:
The ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay utilizes a anti-rat cytokine antibody for immobilization on the microtiter wells and anti-rat cytokine 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 molecules of the cytokine 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 the cytokine is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.
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