Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Intracellular Staining by Flow Cytometry, Neutralization

Cited:

Bioassay, Cell Culture, ELISA Capture, Flow Cytometry, Immunohistochemistry, Immunoprecipitation, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG1 Clone # MP6-XT22

View all TNF-alpha Antibodies »

Product Specifications

Immunogen

mouse TNF-alpha

Specificity

Detects mouse TNF-alpha.

Clonality

Monoclonal

Host

Rat

Isotype

IgG1

Endotoxin Level

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

Scientific Data Images for Mouse TNF-alpha Antibody

Detection of TNF-alpha in Raw264 Mouse Cell Line by Flow Cytometry.

Raw264 mouse macrophage cell line treated with 1 μg/mL LPS overnight was stained with Rat Anti-Mouse TNF-alpha Monoclonal Antibody (Catalog # MAB4101, filled histogram) or isotype control antibody (MAB005, open histogram) followed by Anti-Rat IgG CFS-conjugated Secondary Antibody (F0104). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Kit (FC012). Staining was performed using our Staining Intracellular Molecules protocol.

Detection of TNF-alpha in Mouse Splenocytes by Flow Cytometry.

Mouse splenocytes (A) treated with PMA (50 ng/mL), Ca2+ Ionomycin (200 ng/mL) and Brefeldin A (5 μg/mL) for 4 hours or (B) untreated, were stained with Rat Anti-Mouse TNF-alpha Monoclonal Antibody (Catalog # MAB4101) followed by Anti-Rat IgG PE-conjugated Secondary Antibody (F0105B) and Rat anti-Mouse CD3 APC-conjugated Monoclonal Antibody (FAB4841A). Quadrant markers were set based on isotype control antibody (MAB005). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our protocol for Staining Intracellular Molecules.
Cytotoxicity Induced by TNF-alpha and Neutralization by Mouse TNF-alpha Antibody.

Cytotoxicity Induced by TNF-alpha and Neutralization by Mouse TNF-alpha Antibody.

Recombinant Mouse TNF-a (Catalog # 410-MT) induces cytotoxicity in the the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Mouse TNF-a (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Mouse TNF-a Monoclonal Antibody (Catalog # MAB4101). The ND50 is typically 0.15-0.75 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
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Applications for Mouse TNF-alpha Antibody

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Raw264 cells treated with LPS, fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Kit (Catalog # FC012) and mouse splenocytes treated with PMA and Ca2+ Ionomycin

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 0.15-0.75 µg/mL in the presence of 0.25 ng/mL Recombinant Mouse TNF‑ alpha and 1 µg/mL actinomycin D.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

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.

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-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Mouse TNF-alpha consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD). Within the ECD, mouse TNF-alpha shares 94% aa sequence identity with rat and 70%-77% with bovine, canine, cotton rat, equine, feline, human, porcine, and rhesus 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

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

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

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for Mouse TNF-alpha Antibody

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Mouse TNF-alpha Antibody

For research use only

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