Human MCP-1 ELISA Kit

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.

Target Information

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.

GENE ID 6347
SWISS PROT P13500
SYNONYMS HC11; MCAF; MCP1; MCP-1; SCYA2; GDCF-2; SMC-CF; HSMCR30

Materials Supplied

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

Storage Store at 2 - 8°C

Performance Characteristics

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