Principle of the Assay
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.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
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.
|SYNONYMS||HC11; MCAF; MCP1; MCP-1; SCYA2; GDCF-2; SMC-CF; HSMCR30|
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human MCP-1 (8﹡12)||1 plate|
|Human MCP-1 Standard lyophilized, 1000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human MCP-1 monoclonal antibody||2 vials|
|Streptavidin-HRP solution||2 vials|
|Standard /sample Diluent||1 bottle|
|Biotin-Conjugate antibody Diluent||1 bottle|
|Streptavidin-HRP Diluent||1 bottle|
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 vial|
|Stop Solution||1 vial|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||: The minimum detectable dose was 4pg/mL.|
|ASSAY RANGE||15.60 -1000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human MCP-1. The factors listed below were prepared at 50ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.|
Factors assayed for cross-reactivity
|Recombinant human||GROα, GROβ, GROγ, IL-8, MCP-2, MCP-3, MIP-1α, MIP-1β, RANTES|
|Recombinant mouse||JE/MCP-1, MIP-1α, MIP-1β|
Data Analysis Assistance
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