The viability of L-929 cells, treated with hTNF-α #8902, in the presence of 1 μg/ml actinomycin D was determined. After 24 hour treatment with hTNF-α, viable cells were stained with a tetrazolium salt and the OD450 was determined.
The neutralization ability of Human TNF-α Neutralizing (D1B4) Rabbit mAb on hTNF-α-induced cell cytotoxicity was assessed by adding increasing concentrations of antibody with 1 ng/ml of hTNF-α #8902, before addition to L-929 cells in the presence of 1 μg/ml actinomycin D. After 24 hour incubation, viable cells were stained and % neutralization was calculated.
Neutralizing antibodies can be used to inhibit normal biological function through their binding to biological molecules. These reagents can be used to determine the effects that a particular molecule has in biological systems. TNF-α has known functions of cell cytotoxicity, cell activation, and apoptosis in different cell types. Human TNF-α Neutralizing (D1B4) Rabbit mAb has been shown to neutralize the cytotoxic effects of TNF-α in L-929 mouse fibroblast cells. Utilizing 1 ng/ml of hTNF-α #8902 and 1 µg/ml of actinomycin D, Human TNF-α Neutralizing (D1B4) Rabbit mAb rescued L-929 cells with an ND50 in the range of 4-12 ng/ml.
<0.1 EU/µg of antibody
CST recommends incubation of the neutralizing antibody with intended target for 2 hours at 37ºC before addition to the experiment at an optimal concentration determined by the user.
Lyophilized from a 0.2 µm filtered solution in HEPES with trehalose.
Store lyophilized material at -20ºC. After reconstitution, recommended storage at 4ºC for 1 month or -20ºC for 6 months. Avoid repeated freeze/thawing.
Human TNF-α Neutralizing (D1B4) Rabbit mAb binds to human TNF-α and neutralizes its cytotoxic effects. This antibody does not cross-react with mouse TNF-α.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a recombinant human TNF-α protein.
TNF-α, the prototypical member of the TNF protein superfamily, is a homotrimeric type-II membrane protein (1,2). Membrane-bound TNF-α is cleaved by the metalloprotease TACE/ADAM17 to generate a soluble homotrimer (2). Both membrane and soluble forms of TNF-α are biologically active. TNF-α is produced by a variety of immune cells including T cells, B cells, NK cells, and macrophages (1). Cellular response to TNF-α is mediated through interaction with receptors TNF-R1 and TNF-R2 and results in activation of pathways that favor both cell survival and apoptosis depending on the cell type and biological context. Activation of kinase pathways (including JNK, Erk1/2, p38 MAPK, and NF-κB) promotes the survival of cells, while TNF-α-mediated activation of caspase-8 leads to programmed cell death (1,2). TNF-α plays a key regulatory role in inflammation and host defense against bacterial infection, notably Mycobacterium tuberculosis (3).
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