• Species Reactivity

  Rat

• Specificity

  Detects rat TNF-alpha in direct ELISAs. In ELISAs, this antibody shows less than 3% cross-reactivity with recombinant mouse (rm) TNF‑ alpha and less than 0.2% cross-reactivity with rhTNF‑ alpha, rpTNF‑ alpha, and rhTNF‑ beta.

• Source

  Recombinant Monoclonal Mouse IgG    ₁ Clone # 45418R

• Purification

  Protein A or G purified from cell culture supernatant

• Immunogen

  E. coli-derived recombinant rat TNF-alpha    
  Accession # P16599

• Formulation

  Supplied as a solution in PBS. *Small pack size (SP) is supplied 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

• Rat TNF-alpha Sandwich Immunoassay

  Reagent    

• ELISA Capture (Matched Antibody Pair)

  2-8 µg/mL 

  Rat TNF‑ alpha Antibody (Catalog # )

• ELISA Detection (Matched Antibody Pair)

  0.1-0.4 µg/mL 

  Rat TNF‑ alpha Biotinylated Antibody (Catalog # )

• ELISA Standard

   

  Recombinant Rat TNF-alpha Protein (Catalog # )

• 
  

• 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 (ND     ₅₀) is typically 10-40 µg/mL in the presence of 0.025 ng/mL Recombinant Rat TNF‑ alpha and 0.5 µg/mL Actinomycin D.

Please Note: Optimal dilutions should be determined by each laboratory for each application.  are available in the Technical Information section on our website.

Data Examples

Preparation and Storage

• Shipping

  The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

• 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 opening.

• 6 months, -20 to -70 °C under sterile conditions after opening.

Background: TNF-alpha

Tumor Necrosis Factor Alpha (TNF-alpha ) also known as Cachectin, 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). Rat TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD) (3). Within the ECD, rat TNF-alpha shares 94% aa sequence identity with mouse and 69-76% with bovine, canine, cotton rat, equine, feline, human, porcine, and rhesus macaque 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. Estler, H.C. et al. (1992) Biol. Chem. Hoppe-Seyler 373:271.

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); 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; 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