Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-8/CXCL1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-8/CXCL1 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-8/CXCL1 is added to the wells and binds to the combination of capture antibody-IL-8/CXCL1 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 IL-8/CXCL1 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 IL-8/CXCL1 standard dilutions and IL-8/CXCL1 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Mouse KC, also known as CXCL1 or N51, was originally identified in fibroblasts as a PDGFinduced immediate early gene that encodes a secretory protein of approximately 8 kDa. The protein sequence of mouse KC identified it as a member of the alpha (CXC) chemokine family of inflammatory and immunoregulatory cytokines. Besides mitogen-stimulated fibroblasts, KC expression can be induced in bacterial or LPS-stimulated peritoneal and lung macrophages, endothelial cells and vascular smooth cells. The induction of KC by mitogens has been shown to be inhibited by glucocorticoids.
Mouse KC cDNA encodes a 96 amino acid (aa) residue precursor protein from which the aminoterminal 19 aa residues are cleaved to generate the 77 aa residue mature KC. The protein sequence of mouse KC shows approximately 63% identity to that of mouse MIP-2, another mouse alpha chemokine. In addition, the protein sequence of KC is approximately 60% identical to the human GROs. Like other alpha chemokines, mouse KC is a potent neutrophil attractant and activator. The activities of KC and MIP-2 have been shown to be mediated by the unique mouse IL-8 receptor that shows 71% and 68% aa sequence identity to human IL-8 Rβ and IL-8 Rα, respectively. Since an IL-8 homolog has not been identified in mice, its has been suggested that MIP-2 and KC are the functional homologs of IL-8 and may function as the major proinflammatory alpha chemokines in mice. Increased KC expression has been found to be associated with neutrophil influx in various inflammatory conditions.
|SYNONYMS||KC; Fsp; N51; gro; Gro1; Mgsa; Scyb1|
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to mouse IL-8/CXCL1(8﹡12)||1 plate|
|Mouse IL-8/CXCL1 Standard lyophilized, 1000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-mouse IL-8/CXCL1 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 7.8pg/mL.|
|ASSAY RANGE||15.6-1000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant mouse IL-8/CXCL1. 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 mouse||G-CSF, GM-CSF, IL-1α, IL-1β, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-13, VEGF, LIF, MIP-1, MIP-2, TNF-α, SCF|
|Recombinant human||IL-2, IL-2 sRα, IL-2sRβ|
Data Analysis Assistance
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