Chromogenic solution (TMB) is added and incubated.

Bound enzymelabelled antibody is measured through a chromogenic reaction. After an incubation period allowing the formation of a sandwich: coated M 1 human TFРћВ± M 2, the microtiterplate is washed to remove unbound enzyme labelled antibody. Calibrators and samples react with the capture monoclonal antibody (M 1) coated on microtiter well and with a monoclonal antibody (M 2) labelled with horseradish peroxidase (). The assay uses monoclonal antibodies (Ms) directed against distinct epitopes of TFРћВ±. PRICIPLES OF THE METHOD The DIAsource TFРћВ± EASIA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiterplate. in graft rejection) and have prognostic value (e.g. Besides an insight into pathogenesis, these determinations might provide an aid in diagnosis (e.g. normal high levels of serum TFРћВ± have been described in septic shock, graft rejection, parasitic infections, cancer, post hemofiltrations, during in vivo cytokine (IL2) therapy, etc. TFРћВ± often in combination with other cytokines, has also been involved in several autoimmune diseases and even in the pathogenesis of arteriosclerosis. Clinical application TFРћВ± has a major pathogenic role : in cachexia associated with chronic infectious or cancerous diseases in septic shock where the neutralization of TFРћВ± protects against the associated acute lethality in graft rejection and graftversushost disease and in parasitic infections where TFРћВ± may provide some protection but also favours more severe forms of the disease (e.g. Metabolic activities : TFРћВ± strongly inhibits lipoprotein lipase and adipocyte gene expression. It induces the production of several other inflammatory mediators such as IL1, IL6, colony stimulating factors, prostaglandins, plateletactivating factor (PAF), collagenases, etc. It regulates the production of antibodies by B cells and stimulates cytotoxic T cells. Immunomodulatory and proinflammatory activities : TFРћВ± activates macrophages, neutrophils and eosinophils, as well as endothelial cells (which display procoagulant activity). Conversely, it is angiogenic and stimulates the growth of cultured fibroblasts. The various biological activities of TFРћВ± may be classified as : Antitumoral and growth regulatory activities : TFРћВ± displays a selective toxicity for tumor and virusinfected cells.

Lipopolysaccharide (LPS), the cellwall component of gramnegative bacteria (endotoxin), is a potent stimulus for TFРћВ± production by macrophages and TFРћВ± is an important mediator of the wellknown in vivo effects of LPS such as tumour hemorrhagic necrosis, fever, shock and activation of neutrophils. unglycosylated polypeptide cytokine mainly produced by activated macrophages (monocytes). Biological activities Human Tumor ecrosis Factor Alpha (TFРћВ±) also named cachectin, is a 157 A.A. CLII BACKGROUD For technical assistance or ordering information contact : Tel : +32 () Fax : +32 () A. Rue du Bosquet, 2, B134 Louvainlaeuve, Belgium. Manufactured by : DIAsource ImmunoAssays S.A. Proprietary name : DIAsource TFРћВ± EASIA Kit B. ITEDED USE Immunoenzymetric assay for the in vitro quantitative measurement of human Tumor ecrosis Factor РћВ± (TFРћВ±) in serum. Rue du Bosquet, 2 B134 Louvainlaeuve BelgiumР”Р€ en Read entire protocol before use. 1 TFРћВ± EASIA KAP1751 DIAsource ImmunoAssays S.A.

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