Rat TNF-alpha Antibody

R&D Systems | Catalog # AF-510-NA

R&D Systems
100% Guarantee

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.
Rat TNF-alpha ELISA Standard Curve

Rat TNF-alpha ELISA Standard Curve

Recombinant Rat TNF‑ alpha (Catalog # 510-RT) was serially diluted and captured by Mouse Anti-Rat TNF‑ alpha Monoclonal Antibody (Catalog # MAB510) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-510-NA) 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 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.

=
÷

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

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Rat TNF-alpha Antibody

For research use only

Citations for Rat TNF-alpha Antibody

Customer Reviews for Rat TNF-alpha Antibody

There are currently no reviews for this product. Be the first to review Rat TNF-alpha Antibody and earn rewards!

Have you used Rat TNF-alpha Antibody?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

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

FAQs for Rat TNF-alpha Antibody

Showing  1 - 1 of 1 FAQ Showing All

  • 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. 

Showing  1 - 1 of 1 FAQ Showing All
View all FAQs for Antibodies