Interleukin-10 human ELISA

>Background:Human interleukin-10 (IL-10) is a 19 kDA lymphokine produced by T helper lymphocytes,by monocytes, macrophages and B-lymphocytes. IL-10 was first characterized as a cytokine synthesis inhibitory factor (CSIF) able to inhibit cytokine synthesis by TH1 clones activated in the presence of antigen presenting cells. However, in the absence of monocytes, IL-10 directly inhibits the growth of T-cells triggered by immobilized anti-CD3 MoAb. This proliferation inhibition was found to be a result of specific inhibition of IL-2 production by the responding T-cells. In vitro, Il-10 is a very powerful inhibitor of monokines (including TNF-α, IL-1, IL-6 and IL-8) produced by LPS-activated monocytes and macrophages. The addition of IL-10 to B lymphocytes results in limited cell proliferation but most importantly in very high immunoglobulin production, a result of the transformation of B-cells into plasma cells. Finally, natural killer (NK) cells appear to be another target for the anti-inflammatory properties of IL-10. Indeed, recent data have shown that IL-10 can inhibit antigen induced IFN-γ production by NK-cells by inhibiting not only production but also the stimulatory effects of IL-12 and TNF on IFN-γ production.

Description:
The IL-10-ELISA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiterplate. The assay uses monoclonal antibodies (MAbs) directed against distinct epitopes of IL10. Calibrators and samples react with the capture monoclonal antibody (MAb 1) coated on microtiter well and with a monoclonal antibody (MAb 2) labelled with horseradish peroxidase (HRP). After an incubation period allowing the formation of a sandwich: coated MAb 1 – human IL-10 – MAb 2 – HRP, the microtiterplate is washed to remove unbound enzyme labelled antibody. Bound enzyme-labelled antibody is measured through a chromogenic reaction. Chromogenic solution (TMB) is added and incubated. The reaction is stopped with the addition of Stop Solution and the microtiterplate is then read at the appropriate wavelength. The amount of substrate turnover is determined colourimetrically by measuring the absorbance, which is proportional to the IL-10 concentration. A calibration curve is plotted and IL-10 concentration in samples is determined by interpolation from the calibration curve. The use of the ELISA reader (linearity up to 3 OD units) and a sophisticated data reduction method (polychromatic data reduction) result in a high sensitivity in the low range and in an extended calibration range.

Product features:
The kit contains reagents for 96 determinations;
Microtiter plate consisting of 12x8 (breakapart);
Microtiter plate reader at 450, 490 and 650 nm;
Limit of detection: 1,6 pg/ml;
IVD.