• Species Reactivity

  Porcine

• Specificity

  Detects porcine TNF-alpha in direct ELISAs and Western blots. In Western blots, no cross-reactivity with recombinant human (rh) APRIL, rhFasL, rhGITRL, rhLIGHT, recombinant mouse (rm) TRANCE, rhBAFF, rhTNF-alpha, rmTNF-alpha, recombinant rat TNF-alpha, or rhTNF-beta is observed.

• Source

  Monoclonal Mouse IgG    ₁ Clone # 103302

• Purification

  Protein A or G purified from hybridoma culture supernatant

• Immunogen

  E. coli-derived recombinant porcine TNF-alpha    
  Arg78-Leu232    
  Accession # P23563

• Formulation

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

• Western Blot

  1 µg/mL

  Recombinant Porcine TNF‑ alpha (Catalog # )

• 
  

• Immunocytochemistry

  8-25 µg/mL

  See below

• 
  

• Neutralization

  Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the PK‑15 porcine kidney epithelial 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; Bertoni     et al. (1993) J.  Immunol. Meth.     160:267. The Neutralization Dose (ND    ₅₀) is typically 0.005-0.02 µg/mL in the presence of 0.05 ng/mL Recombinant Porcine TNF‑ alpha and 1 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

• Reconstitution

  Reconstitute at 0.5 mg/mL in sterile PBS.

• Shipping

  The product is shipped at ambient temperature. 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 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). Porcine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 176 aa extracellular domain (ECD) (3). Within the ECD, porcine TNF-alpha shares 69%‑86% aa sequence identity with bovine, canine, cotton rat, equine, feline, human, mouse, 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 costimulation 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. Pauli, U. et al. (1989) Gene 81:185.

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