Mouse IL-6 ELISA Kit

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. Standards and samples are pipetted into the wells and any IL-6 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-6 is added to the wells and 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.

Target Information

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. It is expressed by a variety of normal and transformed lymphoid and nonlymphoid cells.

IL-6 is a prototypic member of the IL-6 superfamily of cytokines that share gp130 as a component required for signal transduction. The mouse, rat and human IL-6 cDNAs have been cloned. The mouse IL-6 cDNA encodes a 211 amino acid (aa) residue precursor polypeptide with a hydrophobic signal sequence that is cleaved to generate the 187 aa residue mature protein. Mouse to rat and human, there is approximately 87% and 39% aa identity, respectively. Although human and mouse IL-6 are equally active on mouse cells, mouse IL-6 is not active on human cells.

The high-affinity IL-6 receptor complex, which mediates IL-6 bioactivity, consists of two membrane glycoproteins. The production of IL-6 is upregulated 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.

GENE ID 16193
SWISS PROT P08505
SYNONYMS Interleukin-6, IL6

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to mouse IL-6(8﹡12) 1 plate
Mouse IL-6 Standard lyophilized, 500 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-mouse IL-6 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 7pg/mL.
ASSAY RANGE 7.8-500 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant mouse 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-6 sR
Recombinant mouse CT-1, gp130, IL-16 sR, IL-11
Recombinant porcine IL-6