Cell Signaling Technology

Product Pathways - NF-kB Signaling

Mouse TNF-α Neutralizing (D2H4) Rabbit mAb #11969

Applications Reactivity Isotype
N M Rabbit IgG

Applications Key:  N=Neutralizing
Reactivity Key:  M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Mouse TNF-α Neutralizing (D2H4) Rabbit mAb binds to mouse TNF-α and neutralizes its cytotoxic effects. This antibody does not cross-react with human TNF-α or human LTA.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant mouse TNF-α protein.

Bioactivity

Bioactivity

The ability of Mouse TNF-α Neutralizing (D2H4) Rabbit mAb to inhibit mTNF-α-induced L-929 cell cytotoxicity was assessed. Cells were incubated with increasing concentrations of antibody in the presence of mTNF-α #5178 (250 pg/ml) and 1 µg/ml actinomycin D. After 24 hr, viable cells were detected by incubation with a tetrazolium salt and the OD450 was determined.

Bioactivity

Bioactivity

The viability of L-929 cells treated with increasing amounts of mTNF-α #5178 in the presence of 1 µg/ml actinomycin D was determined. After a 24 hr treatment, cells were incubated with a tetrazolium salt and the OD450 was determined.

Description

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. Mouse TNF-α Neutralizing (D2H4) Rabbit mAb has been shown to neutralize the cytotoxic effects of TNF-α in L-929 cells in vitro with an ND50 in the range of 1-6 ng/ml.

Endotoxin

<0.1 EU/µg of antibody

Formulation

Lyophilized from a 0.2 µm filtered solution in 10mM HEPES with trehalose.

Background

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, Erk (p44/42), 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). The role of TNF-α in autoimmunity is underscored by research studies that show that blocking TNF-α action may be used to treat rheumatoid arthritis and Crohn’s disease (1,2,4).

  1. Aggarwal, B.B. (2003) Nat Rev Immunol 3, 745-56.
  2. Hehlgans, T. and Pfeffer, K. (2005) Immunology 115, 1-20.
  3. Lin, P.L. et al. (2007) J Investig Dermatol Symp Proc 12, 22-5.
  4. Brennan, F.M. and McInnes, I.B. (2008) J Clin Invest 118, 3537-45.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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