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M Rabbit IgG

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.

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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.

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Product Usage Information

CST recommends incubation of the neutralizing antibody with the intended target for 1 hr at 37ºC before addition to the experiment at an optimal concentration determined by the user.

Application Dilutions
Neutralizing 1:1


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

Storage: Store lyophilized material at -20ºC. After reconstitution, recommended storage at 4ºC for 1 month or -20ºC for 6 months. Avoid repeated freeze/thawing.

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.

Species Reactivity: Mouse

Source / Purification

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

Product 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.


<0.1 EU/µg of antibody

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.

Entrez-Gene Id 21926
Swiss-Prot Acc. P06804

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Mouse TNF-α Neutralizing (D2H4) Rabbit mAb