Monocyte chemoattractant protein-1 (MCP-1) called also monocyte

chemotactic protein-1. MCP-1 is an 8.6kDa protein and belongs to the family of chemotactic cytokines known as Chemokines . MCP-1 is expressed by monocytes, vascular endothelial cells, smooth muscle cells, glomerular mesangial cell, osteoblastic cells, and human pulmonary type-2-like epithelial cells in culture. MCP-1 exhibits chemotactic activity for monocytes/macrophages, basophils, T lymphocytes, particularly memory T cells and natural killer cells and neural stem cells. Elevated levels of MCP-1 are detected during inflammation and immune responses. MCP-1 activity is also important in wound healing as shown by delayed wound repair in MCP-1deficient mice. MCP-1 is implicated in the pathogenesis of disease states that involve monocyte/macrophage infiltrates, such as psoriasis, rheumatoid arthritis and recruitment of monocytes to the arterial wall in atherosclerosis. In addition to its chemotactic function, MCP-1 also induces the expression of IL-10 from macrophages, which favours a TH2 immune response. MCP-1 is a glycopeptide containing one potential N-linked glycosylation site. The extent of MCP-1 glycosylation influences its chemotactic potency and half-life in vivo.

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MCP-1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MCP-1 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for MCP-1 is added to the wells and binds to the combination of capture antibody-MCP-1 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of MCP-1 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven MCP-1 standard dilutions and MCP-1 sample concentration determined.

Figure 1:Schematic diagram of the assay

1. Aluminium pouches with a Microwell Plate coated with antibody to human MCP-1 (812)

2. 2 vials human MCP-1 Standard lyophilized, 1000 pg/ml upon reconstitution

3. 2 vials concentrated Biotin-Conjugate anti-human MCP-1 antibody

4. 2 vials Streptavidin-HRP solution,

5. 1 bottle Standard /sample Diluent

6. 1 bottle Biotin-Conjugate antibody Diluent

7. 1 bottle Streptavidin-HRP Diluent

8. 1 bottle Wash Buffer Concentrate 20x (PBS with 1% Tween-20)

9. 1 vial Substrate Solution

10. 1 vial Stop Solution

11. 4 pieces Adhesive Films

12. package insert

NOTE: [96 Tests]

Unopened Kit：Store at 2 -8° C. Do not use past kit expiration date. opened/ReconstitutedReagents：Please refer to the datasheets for detail information.

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