Cotton Rat TNF-alpha Antibody

Catalog #: MAB10111 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB10111
MAB10111-SP

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Cotton Rat TNF‑ alpha Antibody.

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All TNF-alpha Antibodies

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Cotton Rat TNF-alpha Antibody Summary

Species Reactivity

Cotton Rat

Specificity

Detects cotton rat TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross‑reactivity with recombinant human (rh) TNF-alpha is observed and no cross-reactivity with rhAPRIL, rhBAFF, rhEDA-A2, recombinant mouse (rm) EDA, rhFas Ligand, rhGITR Ligand, rhLIGHT, rhOX40 Ligand, rmTNF-alpha, recombinant porcine TNF-alpha, recombinant rat TNF-alpha, recombinant rhesus macaque TNF-alpha, rhTRAIL, rmTRAIL, rhTRANCE, rhTWEAK, or rhVEGI is observed.

Source

Monoclonal Mouse IgG₁ Clone # 159813

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant cotton rat TNF-alpha
Leu1-Leu156
Accession # AAL18818

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.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

Recombinant Cotton Rat TNF‑ alpha (Catalog # 1011-CR)

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 0.04-0.2 µg/mL in the presence of 1 ng/mL Recombinant Cotton Rat TNF‑ alpha and 1 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

Neutralization

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Cytotoxicity Induced by TNF‑ alpha and Neutralization by Cotton Rat TNF‑ alpha Antibody. Recombinant Cotton Rat TNF-a (Catalog # 1011-CR) 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 Cotton Rat TNF-a (1 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Cotton Rat TNF-a Monoclonal Antibody (Catalog # MAB10111). The ND₅₀ is typically 0.04-0.2 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

Reconstitution Calculator

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 ) 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, apoptosis, and immune system development. TNF-alpha is produced by a wide variety of immune and epithelial cell types (1, 2). The 156 amino acid (aa) cotton rat TNF-alpha is homologous to a portion of the extracellular domain (ECD) of TNF-alpha from other species (3). It shares 64%‑76% aa sequence identity with bovine, canine, equine, feline, human, mouse, porcine, rat, and rhesus TNF-alpha. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer (4). Ligation of this complex induces reverse signaling that promotes lymphocyte co‑stimulation but diminishes monocyte responsiveness (5). Cleavage of membrane bound TNF-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha (6, 7). TNF-alpha trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers (1, 8). TNF-alpha regulates lymphoid tissue development through control of apoptosis (2). It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages (2). TNF-alpha is a key cytokine in the development of several inflammatory disorders (9). It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism (10, 11).

References

Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
Blanco, J.C. et al. (2004) J. Interferon Cytokine Res. 24:21.
Tang, P. et al. (1996) Biochemistry 35:8216.
Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
Black, R.A. et al. (1997) Nature 385:729.
Moss, M.L. et al. (1997) Nature 385:733.
Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
Clark, I.A. (2007) Cytokine Growth Factor Rev. 18:335.
Romanatto, T. et al. (2007) Peptides 28:1050.
Hector, J. et al. (2007) Horm. Metab. Res. 39:250.

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