Key Product Details

Species Reactivity

Validated:

Rat

Cited:

Rat, Transgenic Rat

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant rat TNF-alpha
Leu80-Leu235
Accession # P16599

Specificity

Detects rat TNF‑ alpha in direct ELISAs and Western blots. In Western blots, approximately 25% cross-reactivity with recombinant porcine TNF-alpha is observed and less than 1% cross-reactivity with recombinant human (rh) TNF-beta, rhTNF sRI, and rhTNF sRII is observed. In direct ELISAs, approximately 40% cross-reactivity with recombinant canine TNF-alpha, recombinant feline TNF-alpha, and recombinant rhesus macaque TNF-alpha is observed. Neutralizes the biological activity of rat TNF-alpha. This antibody will also neutralize the biological activity of murine TNF-alpha, but weak neutralization of rhTNF-alpha and no neutralization of rhTNF-beta was observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

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

Scientific Data Images for Rat TNF‑ alpha Antibody

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

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

Recombinant Rat TNF-a (Catalog # 510-RT) induces cytotoxicity in the the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin. Cytotoxicity elicited by Recombinant Rat TNF-a (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-510-NA). The ND50 is typically 0.15-0.75 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
TNF-a antibody in Rat Splenocytes by Immunocytochemistry (ICC).

TNF‑ alpha in Rat Splenocytes.

TNF-a was detected in immersion fixed rat splenocytes using Goat Anti-Rat TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-510-NA) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasmic. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Porcine TNF-alpha by Immunocytochemistry/ Immunofluorescence

Detection of Porcine TNF-alpha by Immunocytochemistry/ Immunofluorescence

Astrocytes activity and GFAP/TNF-alpha double staining in the DG area.Astrocyte activity was measure with GFAP immunofluorescence. Representative photomicrographs show control and LPS-injected rats at different time points up to day 30. (A) Densitometry of GFAP fluorescence intensity was up-regulated from day 1 to day 7. Following LPS injection, astrocytes showed changes in morphology, which was recognized by decreased ramification and hypertrophy of the soma. (B) Double fluorescence staining of GFAP and TNF-alpha in hippocampal astrocytes. The ratio of TNF-alpha positive astrocytes in total astrocytes (GFAP-positive cells) was significantly increased from day 1 and peaking on day 7. Pictures show DG area, data are expressed as mean ± standard error of the mean (n = 4) and compared by 1-way analysis of variance followed with Boferroni post hoc analysis, **p<0.01, ***p<0.001 vs Control. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0106331), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Rat TNF-alpha by Immunocytochemistry/ Immunofluorescence

Detection of Rat TNF-alpha by Immunocytochemistry/ Immunofluorescence

Astrocytes activity and GFAP/TNF-alpha double staining in the DG area.Astrocyte activity was measure with GFAP immunofluorescence. Representative photomicrographs show control and LPS-injected rats at different time points up to day 30. (A) Densitometry of GFAP fluorescence intensity was up-regulated from day 1 to day 7. Following LPS injection, astrocytes showed changes in morphology, which was recognized by decreased ramification and hypertrophy of the soma. (B) Double fluorescence staining of GFAP and TNF-alpha in hippocampal astrocytes. The ratio of TNF-alpha positive astrocytes in total astrocytes (GFAP-positive cells) was significantly increased from day 1 and peaking on day 7. Pictures show DG area, data are expressed as mean ± standard error of the mean (n = 4) and compared by 1-way analysis of variance followed with Boferroni post hoc analysis, **p<0.01, ***p<0.001 vs Control. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0106331), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Rat TNF‑ alpha Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed rat splenocytes

Western Blot

0.1 µg/mL
Sample: Recombinant Rat TNF‑ alpha (Catalog # 510-RT)

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 0.15-0.75 µg/mL in the presence of 0.25 ng/mL Recombinant Rat TNF‑ alpha and 1 µg/mL actinomycin D.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 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). 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 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

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

P16599

Additional TNF-alpha Products

Product Documents for Rat TNF‑ alpha Antibody

Certificate of Analysis

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Product Specific Notices for Rat TNF‑ alpha Antibody

For research use only

Citations for Rat TNF‑ alpha Antibody

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Rat TNF‑ alpha Antibody

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  • Q: What is the expected band size when running Western blot under nonreducing conditions with Catalog # AF-510-NA

    A: Rat TNF-alpha exists as a homotrimer under nonreducing conditions. Each monomer has a molecular weight of ~17 kDa, so the homotrimer form under nonreducing conditions is expected to band at ~51 kDa. 

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