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Soluble Tumor Necrosis Factor Rec II ELISA Kit (GWB-79A646)



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Product Name: Soluble Tumor Necrosis Factor Rec II

Category: Immunology Markers

Sample Type: Serum

Sensitivity: 0,04 ng/mL

Description: Immunoenzymetric assay for the in vitro quantitative measurement of human soluble Tumor Necrosis Factor Receptor N?. 2 (sTNF-RII) in serum

Intended Use: Immunoenzymetric assay for the in vitro quantitative measurement of human soluble Tumor Necrosis Factor Receptor N. 2 (sTNF-RII) in serum.

Background: A. Biological activities
sTNF-RII, also named TBPII or p75, is one of the two receptors of tumor necrosis factors that are present at the surface of many cells. Different processes modulate their presence. IL-2 and the activation of T-lymphocytes increase the presence of both TNF-RI and RII. An increase is also noticed during the maturation of the macrophages or in presence of protein kinases activators. On the opposite, TNF-Rs decrease in the presence of H2O2, epinephrine, Insulin and somatostatin. The two receptors of TNF are able to bind TNF-? and TNF-?. TNF-RII is the form present on monocytes. The gene present in the chromosome 1 codes for a protein of 461 aa. The MW is about 75 kDa ; that suggests an important glycosylation. According to Tartagliata et al, the TNF-RII could be at the origin of the thymocytes and lymphocytes T cytotoxic proliferation. The proliferation of circulating human mononuclear cells under the influence of PHA involves the participation of the two receptors. Soluble forms of the receptors are shedded from the cell membrane and are present in urine, plasma or culture supernatants.

Principles of the method: The GenWay sTNF-RII-EASIA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiterplate. The assay uses monoclonal antibodies (MAbs) directed against distinct epitopes of sTNF-RII. 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 sTNF-RII - 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 sTNF-RII concentration.
A calibration curve is plotted and sTNF-RII 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 and controls 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 WorkingWash 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.
• Alterations in physical appearance of kit reagents may indicate instability or deterioration.

Sensitivity and Specificity: Cross-reactivity and interference were analysed by the addition of different analytes to sTNFRII samples and measuring the apparent sTNF-RII concentration.

Product Name: Soluble Tumor Necrosis Factor Rec II

Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation (By similarity).

Subunit: Homotrimer (By similarity).

Subcellular Location: Cell membrane; Single-pass type II membrane protein (By similarity).

Subcellular Location: Tumor necrosis factor, soluble form: Secreted (By similarity).

Ptm: The soluble form derives from the membrane form by proteolytic processing (By similarity).

Ptm: The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1 (By similarity).

Similarity: Belongs to the tumor necrosis factor family.

Additional Information

Name Soluble Tumor Necrosis Factor Rec II ELISA Kit (GWB-79A646)
Related Product Names Soluble Tumor Necrosis Factor Rec II ELISA Kit; TNF-alpha; Tumor necrosis factor ligand superfamily member 2; TNF-a; Cachectin TNFA; TNFSF2Soluble Tumor Necrosis Factor Rec IIsTNF-RII
Swissprot ID O77764
Molecular Weight 25697
Datasheets/Manuals Printable manual for GWB-79A646
Intended Use Research Use Only