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H Endogenous Rabbit 

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.

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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.

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Product Usage Information

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.

Application Dilutions
Neutralizing 1:1


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

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

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: Human

Source / Purification

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).

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.

Entrez-Gene Id 7124
Swiss-Prot Acc. P01375

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