Rat 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. 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.

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.

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.

GENE ID 24498
SWISS PROT P20607
SYNONYMS ILg6; Ifnb2

Materials Supplied

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

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 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 mouse IL-6
Recombinant rat IL-1α, IL-1β, IL-2, IL-4, IL-10