S7.5 (01) mTNF-
This Mouse TNF-alpha (α) ELISA Kit is to be used for the
quantitativedetermination of mouse tumour necrosis factor alpha (mouse TNF-α) concentrations inserum and cell culture supernatant. This kit is intended FOR LABORATORY RESEARCHUSE ONLY.
Tumor Necrosis Factor alpha (TNF-α) is a multifunctional proinflammatory cytokine,mainly secreted by activated macrophages. The cytokine was named for its remarkableability to cause hemorrhagic necrosis of tumors in mice. It is implicated with a variety of biological procedures including systemic inflammation, cell proliferation, apoptosis, lipidmetabolism, and coagulation. It is well documented that TNF-
functions through itsreceptors, TNFR1 (p55) and TNFR2 (p75).TNF-
plays an important role in the immune response to bacterial, and certain fungal,viral, parasitic invasions, in tissue remodeling, autoimmune-diseases and the necrosis of specific tumors. The pleiotropic effect of TNF-
regulation is attributed to its ability totrigger multiple signaling pathways simultaneously. TNF-
hyper-expression in responseto the components of some bacteria such as LPS can cause life threatening septic shock.Recombinant TNF-α, in combination with chemotherapy, has been applied for synergistictreatment of soft sarcomas, melanomas and other irresectable tumors. Anti-TNF-
therapy has been used for treatment of rheumatoid arthritis.
This mouse TNF-α ELISA is a 3.5 hour solid phase immunoassay readily applicable tomeasure mouse TNF-
levels in serum, plasma, cell culture supernatant in the range of 0 to 1000pg/mL.
PRINCIPLE OF THE ASSAY
This mouse TNF-α enzyme linked immunosorbent assay (ELISA) applies a techniquecalled a quantitative sandwich immunoassay. The microtiter plate provided in this kit hasbeen pre-coated with a monoclonal antibody specific to mouse TNF-α. Standards or samplesare then added to the appropriate microtiter plate wells with a biotin-conjugatedmonoclonal antibody preparation specific for mouse TNF-
and incubated. Mouse TNF-
if present, will bind and become immobilized by the antibody pre-coated on the wells andthen be “sandwiched” by biotin conjugate. The microtiter plate wells are thoroughlywashed to remove unbound mouse TNF-α and other components of the sample. In order to quantify the amount of mouse TNF-α present in the sample, avidin conjugated toHorseradish Peroxidase (HRP) is added to each microplate well and incubated. Avidin is