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7321
Human TNF-α Neutralizing (D1B4) Rabbit mAb
Primary Antibodies

Human TNF-α Neutralizing (D1B4) Rabbit mAb #7321

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Citations (14)

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.

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Isotype Rabbit IgG

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

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. 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.

Endotoxin:

<0.1 EU/µg of antibody

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.

Formulation:

Lyophilized from a 0.2 µm filtered solution in 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

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.

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, 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.

Pathways & Proteins

Explore pathways + proteins related to this product.

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

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