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Mouse TNF-alpha Antibody

R&D Systems, part of Bio-Techne | Catalog # AB-410-NA

R&D Systems, part of Bio-Techne
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AB-410-NA

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse, Transgenic Mouse

Applications

Validated:

Neutralization, Western Blot

Cited:

Cell Culture, Immunohistochemistry, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

E. coli-derived recombinant mouse TNF-alpha
Leu80-Leu235
Accession # P06804

Specificity

Detects mouse TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with human TNF‑ alpha is observed, and less than 5% cross-reactivity with recombinant bovine TNF‑ alpha, recombinant canine TNF‑ alpha, recombinant equine TNF‑ alpha, recombinant feline TNF‑ alpha, and recombinant porcine TNF‑ alpha is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

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

Scientific Data Images for Mouse TNF-alpha Antibody

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), as measured by crystal violet staining. Cytotoxicity elicited by Recombinant Mouse TNF-a (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Mouse TNF-a Polyclonal Antibody (Catalog # AB-410-NA). The ND50 is typically 0.2-0.6 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
Detection of Mouse Mouse TNF-alpha Antibody by Western Blot

Detection of Mouse Mouse TNF-alpha Antibody by Western Blot

Effect of a TNF-alpha -neutralizing antibody on LPS-induced perturbation of muscle differentiation.(A) C2C12 myoblasts were cultured for 48 h in DM alone, LPS (1 μg/mL), or LPS (1 μg/mL) plus TNF-alpha -neutralizing antibody (5 μg/mL). A representative western blot probed with antibodies to myogenin, MyoD, or beta-tubulin (internal standard) is shown. (B and C) Quantification of the data presented in (A). Data are the mean ± SEM of 7–15 independent experiments. (D) C2C12 myoblasts were cultured for 144 h as described in (A). A representative western blot probed with antibodies to MyHC II, myostatin, or beta-tubulin (internal standard) is shown. (E and F) Quantification of the data presented in (D). Data are the mean ± SEM of 5–21 independent experiments. (G) Cells were treated as described in (A), and NF-kappa B activity was analyzed by ELISA. Data are the mean ± SEM of 7 independent experiments performed in duplicate. ***p < 0.001, **p < 0.01, *p < 0.05, ###p < 0.001, ##p < 0.01, #p < 0.05 by one-way ANOVA followed by Tukey’s honest significant difference test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28742154), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Mouse TNF-alpha Antibody by Western Blot

Detection of Mouse Mouse TNF-alpha Antibody by Western Blot

Effect of a TNF-alpha -neutralizing antibody on LPS-induced perturbation of muscle differentiation.(A) C2C12 myoblasts were cultured for 48 h in DM alone, LPS (1 μg/mL), or LPS (1 μg/mL) plus TNF-alpha -neutralizing antibody (5 μg/mL). A representative western blot probed with antibodies to myogenin, MyoD, or beta-tubulin (internal standard) is shown. (B and C) Quantification of the data presented in (A). Data are the mean ± SEM of 7–15 independent experiments. (D) C2C12 myoblasts were cultured for 144 h as described in (A). A representative western blot probed with antibodies to MyHC II, myostatin, or beta-tubulin (internal standard) is shown. (E and F) Quantification of the data presented in (D). Data are the mean ± SEM of 5–21 independent experiments. (G) Cells were treated as described in (A), and NF-kappa B activity was analyzed by ELISA. Data are the mean ± SEM of 7 independent experiments performed in duplicate. ***p < 0.001, **p < 0.01, *p < 0.05, ###p < 0.001, ##p < 0.01, #p < 0.05 by one-way ANOVA followed by Tukey’s honest significant difference test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28742154), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse TNF-alpha Antibody

Application
Recommended Usage

Western Blot

1 µg/mL
Sample: Recombinant Mouse TNF-alpha (Catalog # 410-MT)

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 0.2-0.6 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

Reconstitution

Reconstitute at 1 mg/mL in sterile PBS.

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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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

UniProt

Additional TNF-alpha Products

Product Documents for Mouse TNF-alpha Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse TNF-alpha Antibody

For research use only

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