Rat TNF-alpha Antibody
R&D Systems, part of Bio-Techne | Catalog # AF-510-NA
Scientific images may be resized, cropped, or adjusted for brightness or contrast for presentation purposes. These updates are limited to presentation and do not affect the underlying scientific interpretation of the data. Where available, additional source data may be provided upon request.
Conjugate
Catalog #
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
Product Specifications
Immunogen
E. coli-derived recombinant rat TNF-alpha
Leu80-Leu235
Accession # P16599
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.
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‑ 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
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
Sample: Immersion fixed rat splenocytes
Western Blot
0.1 µg/mL
Sample: Recombinant Rat TNF-alpha (Catalog # 510-RT)
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.
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
- Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
- Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
- Estler, H.C. et al. (1992) Biol. Chem. Hoppe-Seyler 373:271.
- Tang, P. et al. (1996) Biochemistry 35:8216.
- Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
- Black, R.A. et al. (1997) Nature 385:729.
- Moss, M.L. et al. (1997) Nature 385:733.
- Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
- Clark, I.A. (2007) Cytokine Growth Factor Rev. 18:335.
- Romanatto, T. et al. (2007) Peptides 28:1050.
- 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
Gene Symbol
TNF
UniProt
Additional TNF-alpha Products
Product Documents for Rat TNF-alpha Antibody
Product Specific Notices for Rat TNF-alpha Antibody
For research use only
Loading...
Loading...
Loading...
Loading...
Loading...
Loading...