Interleukin-10 (IL-10) is a family of proteins that includes classical members like IL-1α and IL-1β. ILMouse interleukin-10 (IL-10), also known as cytokine synthesis inhibitory factor (CSIF), is a member of the IL-10 alpha-helix cytokine family. This molecular family also includes IL-19, IL-12, IL-22, IL-24, and IL-26/AK155. IL-10 is secreted by many activated hematopoietic stem cells, liver stellate cells, keratinocytes, and placental trophoblasts. IL-10 plays a role in both inhibitory and activating immune regulation. As an anti-inflammatory cytokine, IL-10 suppresses the activation and expansion of Th1 cells and Th17 cells while promoting the formation of M2 macrophages. It has a crucial role in the proliferation of regulatory T cells (Treg) and the expression of B cells. In the tumor microenvironment, IL-10 inhibits the proliferation of regulatory T cells and myeloid-derived suppressor cells. IL-10 plays a significant protective role in limiting tissue damage during joint inflammation and promoting muscle regeneration after injury, but it can also contribute to persistent viral infections.

This assay kit employs the double antibody sandwich enzyme-linked immunosorbent assay (ELISA) technique. Specific anti-mouse IL-10 antibodies with high affinity are pre-coated onto the enzyme-linked plate. In the wells of the enzyme-linked plate, standards, test samples, and biotinylated detection antibodies are added. After incubation, IL-10 present in the samples binds to the solid-phase antibody and the detection antibody, forming an immune complex. After removing unbound substances, streptavidin-horseradish peroxidase labeled with horseradish peroxidase (HRP) is added. After wash step to remove any unbound substances, 3,3′,5,5′-Tetramethylbenzidine (TMB)substrate is added and color develops in proportion to the amount of IL-10 bound initially. The assay is stopped, and the optical density of the wells is determined using a micro-plate reader at 450nm wavelength (with a reference wavelength of 540nm or 570nm) for quantification.

Recovery
Mouse IL-10 of different levels was spiked into cell culture medium samples to determine its recovery rate. The recovery rate ranged from 95-103%, with an average recovery rate of 99%.
Sensitivity
The minimum detectable dose (MDD) of Mouse IL-10 is typically less than 1.8 pg/mL.
The MDD was determined by adding two standard deviations to the mean O.D. value of twenty zero standard replicates and calculating the corresponding concentration.
Linearity
To assess the linearity of the assay, four samples spiked with high concentrations of Mouse IL-10 were diluted with 1×Reagent Diluent to bring them within the detection range.

Specificity
This assay recognizes natural and recombinant Mouse IL-10.
The factors listed below were prepared at 50 ng/mL in 1×Reagent Diluent and assayed for cross-reactivity.
Preparations of the following factors prepared at 50 ng/mL in a mid-range Mouse IL-10 control were assayed for interference. No significant cross-reactivity or interference was observed.