Human TNF-alpha Antibody

Catalog #: MAB2101R Datasheet

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB2101R-100
MAB2101R-SP
MAB2101R-500
MAB2101R-01M

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Human TNF-alpha Antibody Summary

Species Reactivity

Human

Specificity

Detects human TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 25‑50% cross‑reactivity with recombinant porcine TNF‑ alpha and recombinant rhesus macaque TNF-alpha is observed but no cross-reactivity with recombinant cotton rat TNF‑ alpha, recombinant rat
TNF‑ alpha, recombinant human (rh) LT alpha 1/ beta 2, rhLT alpha 2/ beta 1, rhAPRIL, rhBAFF, rhEDA‑A2, recombinant mouse EDA, rhFas Ligand, rhLIGHT, rhOX40 Ligand, rhTRAIL, rhTRANCE, rhTWEAK, or rhVEGI is observed.

Source

Recombinant Monoclonal Mouse IgG₁ Clone # 6401R

Purification

Protein A or G purified from cell culture supernatant

Immunogen

E. coli-derived recombinant human TNF-alpha
Gly57-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

Immunocytochemistry

3-25 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 0.01-0.06 μg/mL in the presence of 0.75 ng/mL Recombinant Human TNF-alpha and 0.5 μg/mL 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

Immunocytochemistry

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TNF‑ alpha in Human PBMCs. TNF-a was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Recombinant Mouse Anti-Human TNF-a Monoclonal Antibody (Catalog # MAB2101R) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

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 L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Human TNF-a (0.75 ng/mL) is neutralized (green line) by increasing concentrations of Recombinant Human TNF-a Monoclonal Antibody (Catalog # MAB2101R). The ND₅₀ is typically 0.01-0.06 µg/mL in the presence of the metabolic inhibitor actinomycin D (0.5 µg/mL).

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