Tumor Necrosis Factor Alpha (TNF-alpha ) also known as Cachectin, 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 macaque 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).
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Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Rat
Cited:
Rat
Applications
Validated:
Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization
Cited:
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Binding Assay, ELISA Development, ELISA Development (Capture), In vivo assay
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # 45418
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Product Specifications
Immunogen
E. coli-derived recombinant rat TNF-alpha
Accession # P16599
Accession # P16599
Specificity
Detects rat TNF-alpha in ELISAs and Western blots. In ELISAs, this antibody shows less than 3% cross-reactivity with recombinant mouse (rm) TNF‑ alpha and less than 0.2% cross-reactivity with rhTNF‑ alpha, rpTNF‑ alpha, and rhTNF‑ beta. In Western blots, this antibody shows 100% cross-reactivity with rmTNF‑ alpha and no cross-reactivity with rfeTNF‑ alpha.
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 Rat TNF-alpha Antibody (45418)
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 crystal violet staining. Cytotoxicity elicited by Recombinant Rat TNF‑a (0.025 ng/mL) is neutralized (green line) by increasing concentrations of Rat TNF‑a Monoclonal Antibody (Catalog # MAB510). The ND50 is typically 10-40 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
Detection of Rat TNF-alpha by Block/Neutralize
IL-10 receptors expressed on hippocampal neurons.(A) Expression of IL-10 receptor mRNAs in hippocampal neurons. The expression of the IL-10 receptor was identified using RT-PCR. The mRNAs of IL-10 receptor alpha and beta were expressed in the hippocampal neurons. IL-10 receptor alpha was expressed mainly in hippocampal neurons of DIV 7. (1, cultured hippocampal neurons at DIV 7; 2, cultured hippocampal neurons at DIV 15; 3, mixed glial culture at DIV 7; 4, mixed glial culture at DIV 15; 5, microglia) Quantification (DIV 15 neuron/ DIV 7 neuron): IL-10 receptor alpha, 0.61; IL-10 receptor beta, 1.06. (B) Expression of IL-10 receptor proteins in cultured hippocampal neurons. Similar to the expression of mRNA, the IL-10 receptor alpha protein was expressed mainly in neurons of DIV 7. Anti-IL-10 receptor alpha antibodies (0.5 μg/ml, Santa Cruz, sc-985) were used for western blotting [27]. Quantification (DIV 15 neuron/ DIV 7 neurons): IL-10 receptor alpha, 0.73.(C) Expression of IL-10 receptor proteins in the developing rat brains. The IL-10 receptor alpha proteins were expressed mainly in the developing brains of embryonic and early postnatal days (E18~P3). Quantification of IL-10 receptor alpha : E18, 0.30; P1, 0.27; P3, 1.0; P7, 0.22; P14, 0.20; P3W, 0.17; P6W, 0.15 (E, embryonic days; P, postnatal days).(D) Images of IL-10 receptor expressions in cultured hippocampal neurons. Hippocampal neurons of DIV 6 were stained with antibodies of IL-10 receptor alpha (5 μg/ml, Santa Cruz, sc-985) (red) and MAP2 (the neuronal marker, green) after treatment with 4% formaldehyde and then -20 °C methanol. Hippocampal neurons expressed IL-10 receptor proteins comparable to spinal neurons or cortical neurons.(E) The induction of synaptic formation by microglia was antagonized by the neutralizing antibody of IL-10 receptor alpha. When anti-mouse IL-10 receptor alpha antibody was applied to the co-culture of mouse microglia and mouse hippocampal neurons, the density of dendritic spines was significantly decreased compared with the con
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 (45418)
Application
Recommended Usage
Western Blot
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 10-40 µg/mL in the presence of 0.025 ng/mL Recombinant Rat TNF‑ alpha.
Rat TNF-alpha Sandwich Immunoassay
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 2 reviews rated 5 using MAB510 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.
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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
Background: TNF-alpha
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 (45418)
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.
Product Specific Notices for Rat TNF-alpha Antibody (45418)
For research use only
Related Research Areas
Citations for Rat TNF-alpha Antibody (45418)
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Customer Reviews for Rat TNF-alpha Antibody (45418) (2)
5 out of 5
2 Customer Ratings
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Application: Western BlotSample Tested: SerumSpecies: RatVerified Customer | Posted 10/18/2021
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Application: ELISASample Tested: Serum and PlasmaSpecies: RatVerified Customer | Posted 11/08/2017An ELISA was build targeting rat TNFa using this antibody as a capture and BAF510 as the detection. The assay was used to measure TNFa in rat serum and plasma samples.
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
IL-15 Signaling Pathways and their Primary Biological Effects in Different Immune Cell Types
Innate Lymphoid Cell Differentiation Pathways
mTOR Signaling Pathway
NOD-like Receptor Signaling Pathways
Th1 Differentiation Pathway
TNF Superfamily Pathway: Human Ligand-Receptor Interactions & their Associated Functions
Toll-Like Receptor Signaling Pathways
