Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-6 has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps, any IL-6 present is bound by the immobilized antibody and the detection antibody specific for IL-6 is binds to the combination of capture antibody-IL-6 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-6 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-6 standard dilutions and IL-6 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interleukin 6 (IL-6) is a multifunctional cytokine that plays important roles in host defense, acute phase reactions, immune responses, nerve cell functions and hematopoiesis.
IL-6 is a prototypic member of the IL-6 superfamily of cytokines that share gp130 as a component required for signal transduction. The rat, mouse and human IL-6 cDNAs have been cloned. Rat IL-6 cDNA encodes a 211 amino acid (aa) residue precursor polypeptide with a hydrophobic signal peptide that is cleaved to generate the 187 aa residue mature protein. At the protein sequence level, there is approximately 39% identity between rat and human, and 87% identity between mouse and rat IL-6.
IL-6 is expressed by a variety of normal and transformed lymphoid and non-lymphoid cells. The production of IL-6 is up-regulated by numerous signals such as mitogenic or antigenic stimulation, lipopolysaccharides, calcium ionophores, cytokines and viruses. IL-4, IL-10 and IL-13 inhibit IL-6 expression in monocytes. Elevated serum IL-6 levels have been observed in a number of pathological conditions, including bacterial and viral infections, trauma, autoimmune diseases, inflammations and malignancies.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with antibody to rat IL-6 (8-12)||1 plate|
|Rat IL-6 Standard lyophilized, 4000pg/ml upon reconstitution||2 vials|
|concentrated Biotin-Conjugate anti-rat IL-6 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 32pg/mL.|
|ASSAY RANGE||62.50 - 4000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant rat IL-6. 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||IL-6, IL-6sR|
|Recombinant rat||IL-1α, IL-1β, IL-2, IL-4, IL-10|
Data Analysis Assistance
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