Porcine TNF-alpha Antibody (103302)

R&D Systems | Catalog # MAB6903

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Porcine

Cited:

Porcine

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Western Blot, Flow Cytometry, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 103302
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

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

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

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

Scientific Data Images for Porcine TNF-alpha Antibody (103302)

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Porcine TNF‑ alpha Antibody.

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Porcine TNF‑ alpha Antibody.

Recombinant Porcine TNF-a (Catalog # 690-PT) induces cytotoxicity in the the PK-15 porcine kidney epithelial cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Porcine TNF-a (0.05 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Porcine TNF-a Monoclonal Antibody (Catalog # MAB6903). The ND50 is typically 0.005-0.02 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
TNF-a antibody in Porcine PBMCs by Immunocytochemistry (ICC).

TNF‑ alpha in Porcine PBMCs.

TNF-a was detected in immersion fixed porcine peripheral blood mononuclear cells treated with calcium ionomycin and PMA using Mouse Anti-Porcine TNF-a Monoclonal Antibody (Catalog # MAB6903) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Porcine TNF-alpha by Western Blot

Detection of Porcine TNF-alpha by Western Blot

Protein expression of TNF-alpha and IL-6 were detected by Western blot. (A) Sham-operation group; (B) CME group; (C) treatment group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25130514), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Porcine TNF-alpha by Immunohistochemistry-Paraffin

Detection of Porcine TNF-alpha by Immunohistochemistry-Paraffin

Protein expression of TNF-alpha and IL-6 were detected by immunohistochemistry (×400). (A) Sham-operation group; (B) CME group; (C) treatment group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25130514), licensed under a CC-BY license. Not internally tested by R&D Systems.
Porcine TNF-alpha ELISA Standard Curve

Porcine TNF-alpha ELISA Standard Curve

Recombinant Porcine TNF‑ alpha (Catalog # 690-PT) was serially diluted and captured by Mouse Anti-Porcine TNF‑ alpha Monoclonal Antibody (Catalog # MAB6902) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Porcine TNF‑ alpha Monoclonal Antibody (Catalog # MAB6903) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Porcine TNF-alpha Antibody (103302)

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed porcine peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA

Western Blot

1 µg/mL
Sample: Recombinant Porcine TNF‑ alpha (Catalog # 690-PT)

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 (ND50) 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.

Reviewed Applications

Read 1 review rated 4 using MAB6903 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

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

P23563

Additional TNF-alpha Products

Product Documents for Porcine TNF-alpha Antibody (103302)

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Product Specific Notices for Porcine TNF-alpha Antibody (103302)

For research use only

Citations for Porcine TNF-alpha Antibody (103302)

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  • Porcine TNF-alpha Antibody
    Name: Leen Hermans
    Application: Block/Neutralize
    Sample Tested: Cell Culture Samples
    Species: Porcine
    Verified Customer | Posted 10/08/2020
    Porcine TNF-alpha Antibody (103302) MAB6903

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