Interleukin-10 Kit

Brand

GENWAY

Overview

Intended Use: Immunoenzymetric assay for the in vitro quantita t ive measurement of human interleukin-10 (IL-10) in serum. Background: A. Biological activitiesHuman interleukin-10 (IL-10) is a 19 kDA lymphokine produced by T helper lymphocytes by monocytes macrophages and B-lymphocytes. IL-10 was ¬rst 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 byimmobilized anti-CD3 MoAb. This proliferation inhibition was found to be a result of speci¬c inhibition of IL-2 production by the responding T- cells. In vitro Il-10 is a very powerful inhibitor of monokines (including TNF- ? 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-in ammatory properties of IL-10. Indeed recent data have shown that IL-10 can inhibit antigen induced IFN? -production by NK- cells by inhibiting not only production but also the stimulatory e ects of IL-12 and TNF on IFN-? production. Principles of the method: The IL-10-EASIA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiterplate. The assay uses monoclonal antibodies (MAbs) directed against distinct epitopes of IL-10. 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 enzymelabelled 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 EASIA 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. Storage and Stability: ? Before opening or reconstitution all kits components are stable until the expiry date indicated on the vial label if kept at 2 to 8C. ? Unused strips must be stored at 2-8C in a sealed bag containing a desiccant until expiration date. ? After reconstitution calibrators controls and Specimen Diluent are stable for 4 days at 2 to 8C. For longer storage periods aliquots should be made and kept at -20C for maximum 2 months. Avoid successive freeze thaw cycles. ? The concentrated Wash Solution is stable at room temperature until expiration date. ? Freshly prepared Working Wash solution should be used on the same day. ? After its first use the conjugate is stable until expiry date if kept in the original well-closed vial at 2 to 8C. ? The freshly prepared revelation solution is stable before use for maximum 15 minutes at room temperature and must be discarded afterwards. ? Alterations in physical appearance of kit reagents may indicate instability or deterioration. Sensitivity and Specificity: No significant cross-reaction was observed in presence of 50 ng of IL-1? IL-1? IL-1ra IL-2 IL-3 IL-4 IL-6 IL-7 IL-8 TNF-? TNF-? IFN-? IFN-? IFN-? TGF-? GM-CSF OSM MIP-1? MIP-1? LIF MCP-1 G-CSF RANTES PF-4 ?TG GRO IP-10 and SCF. This IL-10 assay is specific for human natural and recombinant IL-10. A very low level (

spezie

other

detection target

Interleukin-10

pack size

96 tests