Rat IL-10 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-10 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-10 present is bound by the immobilized antibody and the detection antibody specific for IL-10 is binds to the combination of capture antibody-IL-10 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-10 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-10 standard dilutions and IL-10 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-10 (IL-10), also known as cytokine synthesis inhibitory factor (CSIF), is the charter member of the IL-10 α-helical cytokine family that also includes IL-19, IL-20, IL-22, IL-24, and IL-26/AK155. IL-10 is a pleiotropic cytokine that exerts immunosuppressive or immunostimulatory effects in a variety of cell types. In 1989, Fiorentino et al. found that the mouse Th2 cell line D10.G4.1 produced a new cytokine that inhibits the transcription of cytokine mRNA in Th1 cell line, called cytokine synthesis inhibitory factor (CSIF) and named IL-10 in the same year.

Mouse, rat and human IL-10 cDNA sequences each contain 178 amino acid residues and have an 18 amino acid signal peptide sequence. The mature IL-10 molecule is 160 amino acid residues, and the mouse and human IL-10 molecules contain 5 and 4 cysteine residues respectively. Their molecular weight ranges from 35-40 kDa. At the amino acid level, rat IL-10 has 85% and 74% homology to human and mouse IL-10 respectively. Rat and human IL-10 act on mouse-derived cells, whereas mouse IL-10 does not cross-react to human cells.

IL-10 mediates its biological activities through a heteromeric receptor complex composed of the type II cytokine receptor subunits IL-10 Rα and IL-10 Rβ. IL-10 Rα is a 110 kDa transmembrane glycoprotein that is expressed on lymphocytes, NK cells, macrophages, monocytes, astrocytes, intestinal epithelial cells, cytotrophoblasts, and activated hepatic stellate cells, while the 75 kDa transmembrane IL-10 Rβ is widely expressed. The IL-10 dimer binds to two IL-10 Rα chains, triggering recruitment of two IL-10 Rβ chains. IL-10 Rβ does not bind IL-10 directly but is required for signal transduction. IL-10 Rβ also associates with IL-20 Rα, IL-22 Rα1, or IL-28 Rα to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29.

Antagonists of IL-10 may have anti-EB virus effects; IL-10 may become an anti-inflammatory treatment by promoting the expression of IL-1 Rα by monocytes. Animal experiments show that IL-10 can effectively prevent the death of mouse shock induced by LPS.

GENE ID 25325
SWISS PROT P29456
SYNONYMS IL10X

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with antibody to rat IL-10 (8-12) 1 plate
Rat IL-10 standard lyophilized, 2000pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-rat IL-10 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 15pg/mL.
ASSAY RANGE 31.25 - 2000 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant Rat IL-10. 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 rat CINC-1, GDNF, GM-CSF, IFN-γ, β-NGF, PDGF-BB, TNF-α, IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-18
Recombinant human IL-10, IL-10sR
Recombinant mouse IL-10 sR

 

Data Analysis Assistance

We have partnered with MyAssays to offer you an easy to use and versatile tool to analyze the data you receive using our ELISA Kit. Click the link below to be directed to the data analysis tool provided by MyAssays specifically for BSKR1014.

https://www.myassays.com/bioss-rat-il-10-elisa-kit.assay