Mouse IL-27 p28 ELISA Kit

Sample

cell culture supernatants, serum, plasma

Species Reactivity

Mouse

Intended Use

For the quantitative determination of mouse Interleukin 27 p28 subunit (IL-27 p28) concentrations in cell culture supernates, serum, and plasma.

Contents of Kit

1. Mouse IL-27 p28 Microplate
2. Mouse IL-27 p28 Conjugate
3. Mouse IL-27 p28 Standard
4. Mouse IL-27 p28 Control
5. Assay Diluent
6. Calibrator Diluent
7. Wash Buffer Concentrate
8. Color Reagent A
9. Color Reagent B
10. Stop Solution

Storage

Store the unopened kit at 2-8 °C. Do not use past kit expiration date.

Precision

##Intra-assay Precision## (Precision within an assay)
Three samples of known concentration were tested twenty times on one plate to assess intraassay precision.
##Inter-assay Precision## (Precision between assays)
Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of kit components.

Sensitivity

Eighteen assays were evaluated and the minimum detectable dose (MDD) of mouse IL-27 p28 ranged from 0.3-4.7 pg/mL. The mean MDD was 1.5 pg/mL.
The MDD was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

General Description

Interleukin 27 (IL-27) is a non-covalently linked heterodimeric cytokine that is structurally related to IL-12 (1). It is composed of an Epstein-Barr virus-induced molecule 3 (EBI3) subunit linked with the IL-27 p28 subunit (2-4). IL-27 p28, a 28 kDa glycoprotein belonging to the type I cytokine family, is homologous to IL-12 p35, IL-23 p19, and IL-6 (5). EBI3, a 34 kDa glycoprotein containing two fibronectin type III domains, belongs to the type I cytokine receptor family. It is homologous to the p40 subunit of IL-12 and IL-23 and to the extracellular domain of IL-6 Rα (6). In addition to IL-27 p28, EBI3 heterodimerizes with IL-12 p35 and can also exist as a homodimer (7). IL-27 is produced by activated antigen presenting cells including monocytes, endothelial cells, and dendritic cells (8). Its expression is induced in response to various inflammatory stimuli (7, 8).
IL-27 binds and signals through a heterodimeric receptor complex consisting of WSX-1 (TCCR) and gp130, both belonging to the cytokine receptor superfamily (9, 10). WSX-1 is specific for IL-27 and is expressed on resting/naive CD4+ T cells, CD8+ T cells, NK cells, dendritic cells, monocytes, mast cells, and B cells (5, 9, 10). In contrast, gp130 is ubiquitously expressed by a variety of immune and non-immune cells and functions as a subunit of the receptor complexes for at least seven other cytokines (5). IL-27 has both pro- and anti-inflammatory properties (11). In response to infection, IL-27 induces monocytes and mast cells to secrete pro-inflammatory cytokines. It induces naive CD4+ T cells to proliferate and develop Th1 cell responses. IL-27 also promotes effector functions of NK cells and CD8+ T cells. As an anti-inflammatory immunomodulator, IL-27 has been found to have the ability to inhibit Th1 or Th2 responses and restrict the strength and duration of adaptive immune responses. The Quantikine Mouse IL-27 p28/IL-30 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IL-27 p28 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse IL-27 p28 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor.
Results obtained using natural mouse IL-27 p28 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse IL-27 p28.

Standard Curve