Human TNF-alpha Antibody

Catalog #: MAB2103
1 Citation Datasheet

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB2103-100
MAB2103-SP

Detection of Human TNF‑ alpha by Western Blot.

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All TNF-alpha Antibodies

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Citations (1)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human TNF-alpha Antibody Summary

Species Reactivity

Human

Specificity

Detects human TNF‑ alpha in direct ELISAs.

Source

Recombinant Monoclonal Rabbit IgG Clone # 2342C

Purification

Protein A or G purified from cell culture supernatant

Immunogen

E. coli-derived recombinant human TNF‑ alpha
Val77-Leu233
Accession # P01375

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

See below

Neutralization

Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. Matthews, N. and M.L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 221. The Neutralization Dose (ND₅₀) is typically 1-10 ng/mL in the presence of 0.25 ng/mL Recombinant Human TNF‑ alpha and actinomycin D.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human TNF‑ alpha by Western Blot. Western blot shows lysates of CHO Chinese hamster ovary cell line either mock transfected or transfected with human TNF- alpha. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human TNF‑ alpha Monoclonal Antibody (Catalog # MAB2103) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Specific bands were detected for TNF‑ alpha at approximately 13 and 16 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Neutralization

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Cytotoxicity Induced by TNF-alpha and Neutralization by Human TNF-alpha Antibody. Recombinant Human TNF-a (Catalog # 210-TA) induces cytotoxicity in the the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Cytotoxicity elicited by Recombinant Human TNF-a (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human TNF-a Monoclonal Antibody (Catalog # MAB2103). The ND₅₀ is typically 1-10 ng/mL in the presence of the metabolic inhibitor actinomycin D.

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-alpha, TNF- alpha, TNFA ), also known as Cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism. TNF- is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Human TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-alpha shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface. Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa molecular weight soluble trimer of the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 80 kDa TNF RII, both of which are also expressed as homotrimers present on virtually all cell types. Both type I and type II receptors bind TNF-alpha with comparable affinity, although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha.

Long Name

Tumor Necrosis Factor alpha

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Alternate Names

APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

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Citation for Human TNF-alpha Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

Tissue-regenerative potential of the secretome of gamma-irradiated peripheral blood mononuclear cells is mediated via TNFRSF1B-induced necroptosis
Authors: E Simader, L Beer, M Laggner, V Vorstandle, A Gugerell, M Erb, P Kalinina, D Copic, D Moser, A Spittler, E Tschachler, H Jan Ankers, M Mildner
Cell Death Dis, 2019-09-30;10(10):729.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot