Cotton Rat TNF-alpha Antibody

Catalog # Availability Size / Price Qty
MAB1011
MAB1011-SP
Cytotoxicity Induced by TNF‑ alpha  and Neutralization by Cotton Rat TNF‑ alpha  Antibody.
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Cotton Rat TNF-alpha Antibody Summary

Species Reactivity
Cotton Rat
Specificity
Detects recombinant cotton rat TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant human (rh) EDA-A2 is observed, 5% cross-reactivity with rhTRAIL and recombinant mouse (rm) TRAIL is observed, and no cross‑reactivity with rhTNF-alpha, rmTNF-alpha, rhAPRIL, rhBAFF, rhEDA-A2, rhGITR Ligand, rhLIGHT, rhOX40 Ligand, rhTWEAK, rhVEGI, or rmTRANCE is observed.
Source
Monoclonal Mouse IgG2B Clone # 159811
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.
Label
Unconjugated

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 0.05- 0.3 µg/mL in the presence of 0.5 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 Cytotoxicity Induced by TNF‑ alpha  and Neutralization by Cotton Rat TNF‑ alpha  Antibody. View Larger

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 Resazurin (Catalog # AR002). Cytotoxicity elicited by Recombinant Cotton Rat TNF-a (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Cotton Rat TNF-a Monoclonal Antibody (Catalog # MAB1011). The ND50 is typically 0.05- 0.3 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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
  1. Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
  2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
  3. Blanco, J.C. et al. (2004) J. Interferon Cytokine Res. 24:21.
  4. Tang, P. et al. (1996) Biochemistry 35:8216.
  5. Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
  6. Black, R.A. et al. (1997) Nature 385:729.
  7. Moss, M.L. et al. (1997) Nature 385:733.
  8. Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
  9. Clark, I.A. (2007) Cytokine Growth Factor Rev. 18:335.
  10. Romanatto, T. et al. (2007) Peptides 28:1050.
  11. 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

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