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  • Product Name
    TNFA Antibody
  • Catalog No.
    AF7014
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity:
    Human, Mouse, Rat
  • Prediction:
    Pig(100%), Bovine(90%), Sheep(90%), Dog(90%)
  • UniProt
  • Mol.Wt.
    17/30-35kDa
  • Concentration
    1mg/ml
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Product Information

Alternative Names:Expand▼

APC1; APC1 protein; Cachectin; DIF; Differentiation inducing factor; Macrophage cytotoxic factor; Tnf; TNF superfamily, member 2; TNF, macrophage derived; TNF, monocyte derived; TNF-a; TNF-alpha; TNFA; TNFA_HUMAN; TNFSF2; Tumor necrosis factor alpha; Tumor necrosis factor; Tumor necrosis factor ligand superfamily member 2; Tumor Necrosis Factor, Membrane Form; Tumor necrosis factor, soluble form;

Applications:

WB 1:500, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(100%), Bovine(90%), Sheep(90%), Dog(90%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Specificity:

TNFA Antibody detects endogenous levels of total TNFA.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

PBS, pH 7.4,50% glycerol.

Immunogen Information

Immunogen:

A synthesized peptide derived from human TNFA, corresponding to a region within the internal amino acids.

Uniprot:



>>Visit The Human Protein Atlas

Gene id:

Molecular Weight:

Observed Mol.Wt.: 17/30-35kDa.
Predicted Mol.Wt.: 26kDa.

Subcellular Location:

Secreted and Cell membrane.

Description:

This gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation.

Sequence:
        10         20         30         40         50
MSTESMIRDV ELAEEALPKK TGGPQGSRRC LFLSLFSFLI VAGATTLFCL
60 70 80 90 100
LHFGVIGPQR EEFPRDLSLI SPLAQAVRSS SRTPSDKPVA HVVANPQAEG
110 120 130 140 150
QLQWLNRRAN ALLANGVELR DNQLVVPSEG LYLIYSQVLF KGQGCPSTHV
160 170 180 190 200
LLTHTISRIA VSYQTKVNLL SAIKSPCQRE TPEGAEAKPW YEPIYLGGVF
210 220 230
QLEKGDRLSA EINRPDYLDF AESGQVYFGI IAL

Background

Function:

Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Impairs regulatory T-cells (Treg) function in individuals with rheumatoid arthritis via FOXP3 dephosphorylation. Upregulates the expression of protein phosphatase 1 (PP1), which dephosphorylates the key 'Ser-418' residue of FOXP3, thereby inactivating FOXP3 and rendering Treg cells functionally defective (PubMed:23396208). Key mediator of cell death in the anticancer action of BCG-stimulated neutrophils in combination with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line (PubMed:22517918).

Post-translational Modifications:

The soluble form derives from the membrane form by proteolytic processing. The membrane-bound form is further proteolytically processed by SPPL2A or SPPL2B through regulated intramembrane proteolysis producing TNF intracellular domains (ICD1 and ICD2) released in the cytosol and TNF C-domain 1 and C-domain 2 secreted into the extracellular space.The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.

Subcellular Location:

Plasma membrane;Extracellular region or secreted;Extracellular region or secreted;Extracellular region or secreted;

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Subunit Structure:

Homotrimer. Interacts with SPPL2B.

Similarity:

Belongs to the tumor necrosis factor family.

Research Fields

Research Fields:

· Cellular Processes > Cell growth and death > Apoptosis.(View pathway)
· Cellular Processes > Cell growth and death > Necroptosis.(View pathway)
· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > TNF signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > MAPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > mTOR signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.(View pathway)
· Environmental Information Processing > Signaling molecules and interaction > Cytokine-cytokine receptor interaction.(View pathway)
· Human Diseases > Immune diseases > Graft-versus-host disease.
· Human Diseases > Immune diseases > Systemic lupus erythematosus.
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Immune diseases > Allograft rejection.
· Human Diseases > Endocrine and metabolic diseases > Type II diabetes mellitus.
· Human Diseases > Cardiovascular diseases > Dilated cardiomyopathy (DCM).
· Human Diseases > Endocrine and metabolic diseases > Type I diabetes mellitus.
· Human Diseases > Immune diseases > Rheumatoid arthritis.
· Human Diseases > Cardiovascular diseases > Hypertrophic cardiomyopathy (HCM).
· Human Diseases > Infectious diseases: Bacterial > Legionellosis.
· Human Diseases > Infectious diseases: Bacterial > Pertussis.
· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).
· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.
· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.
· Human Diseases > Infectious diseases: Viral > Hepatitis C.
· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).
· Human Diseases > Infectious diseases: Parasitic > African trypanosomiasis.
· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.
· Human Diseases > Infectious diseases: Viral > Influenza A.
· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).
· Human Diseases > Infectious diseases: Parasitic > Malaria.
· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.
· Human Diseases > Infectious diseases: Viral > HTLV-I infection.
· Human Diseases > Drug resistance: Antineoplastic > Antifolate resistance.
· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).
· Human Diseases > Immune diseases > Asthma.
· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.
· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.
· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > Antigen processing and presentation.(View pathway)
· Organismal Systems > Immune system > Hematopoietic cell lineage.(View pathway)
· Organismal Systems > Immune system > Toll-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > IL-17 signaling pathway.(View pathway)
· Organismal Systems > Immune system > T cell receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > Fc epsilon RI signaling pathway.(View pathway)
· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity.(View pathway)
· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.
· Organismal Systems > Development > Osteoclast differentiation.(View pathway)
· Organismal Systems > Immune system > NOD-like receptor signaling pathway.(View pathway)

Western blot analysis of extracts from various samples, using TNFA Antibody. Lane 1: UV treated MCF7 cells, blocked with antigen-specific peptides, Lane 2: UV treated MCF7 cells, Lane 3: Serum starvation treated 293, Lane 4: H2O2 treated HepG2 cells.
Western blot analysis of extracts from rat brain, using TNFA Antibody. The lane on the right was treated with blocking peptide.
Immunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue, using TNFA antibody.
AF7014 staining A-431 cells by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25°C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37°C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) antibody(Red), diluted at 1/600, was used as secondary antibody.

Reference Citations:

1). Zheng ZW et al. Development of an Accurate and Proactive Immunomodulatory Strategy to Improve Bone Substitute Material-Mediated Osteogenesis and Angiogenesis. Theranostics 2018 Oct 29;8(19):5482-5500 (PubMed: 30555559) [IF=8.063]

2). Zhang DD et al. A novel IL-1RA-PEP fusion protein with enhanced brain penetration ameliorates cerebral ischemia-reperfusion injury by inhibition of oxidative stress and neuroinflammation. Exp Neurol 2017 Nov;297:1-13 (PubMed: 28602833) [IF=4.562]

Application: IHC    Species:mouse;    Sample:Not available

The expression levels of IL-6 and TNF-α in the cerebral tissues of vehicle group were increased significantly in comparison with that of the control group.


3). Wu M et al. Resveratrol delays polycystic kidney disease progression through attenuation of nuclear factor κB-induced inflammation. Nephrol Dial Transplant 2016 Nov;31(11):1826-1834 (PubMed: 27190325) [IF=4.198]

Application: WB    Species:rat;    Sample:Not available

FIGURE 3: Pro-inflammatory factors, anti-oxidant enzyme and cell signaling pathways in resveratrol (Res)- or vehicle-treated cystic kidneys. (A) TNF-α, MCP-1, CFB and SOD2 were analyzed by western blot in 9-week-old +/+ and Cy/+ kidneys. (B–E) Immunohistochemical staining for oxidative stress markers 8-OHdG and nitrotyrosine in the tubulointerstitial area. Computer-assisted morphometry was used to quantify changes of 8-OHdG and nitrotyrosine in each group. Scale bar = 50 µm. (F) NF-κB pathway ( p-p65, p65, p105 and p50) and mTOR pathway ( p-S6K and total S6K) were analyzed by western blot in 9-week-old +/+ and Cy/+ kidneys. Blots are representative of three independent experiments.


4). Hu J et al. Effects of Xin-Ji-Er-Kang on heart failure induced by myocardial infarction: Role of inflammation, oxidative stress and endothelial dysfunction. Phytomedicine 2018 Mar 15;42:245-257 (PubMed: 29655692) [IF=4.180]

5). Hu J et al. Effects of Xin-Ji-Er-Kang on heart failure induced by myocardial infarction: Role of inflammation, oxidative stress and endothelial dysfunction. Phytomedicine 2018 Mar 15;42:245-257 (PubMed: 29655692) [IF=4.180]

6). Sheng X et al. Knockdown the P2X3 receptor in the stellate ganglia of rats relieved the diabetic cardiac autonomic neuropathy. Neurochem Int 2018 Nov;120:206-212 (PubMed: 30196147) [IF=3.994]

7). Feng Y et al. Neuroprotective Effects of Resatorvid Against Traumatic Brain Injury in Rat: Involvement of Neuronal Autophagy and TLR4 Signaling Pathway. Cell Mol Neurobiol 2017 Jan;37(1):155-168 (PubMed: 26961544) [IF=3.811]

Application: WB    Species:rat;    Sample:rat hippocampus

Fig. 7 a Western blot analysis demonstrates levels of TNF-a and IL-1b in the hippocampus of rats at 12, 24, and 48 h following TBI or sham operation (n = 5/group). b Densitometry of the TNF-a and IL-1b band correlated to the b-actin band. The bars represent the mean ± SE (n = 5/group). The results demonstrated that a significant increase of TNF-a and IL-1b expression in the TBI group (*p\0.01 vs. sham group), and treatment with TAK-242 significantly downregulated the level of TNF-a and IL-1b protein expression at 12, 24, and 48 h following TBI (# p\0.05 vs. TBI group). TNF-a, tumor necrosis factor-a; IL-1b, interleukin-1b


8). Zeng SY et al. Nox1/4 dual inhibitor GKT137831 attenuates hypertensive cardiac remodelling associating with the inhibition of ADAM17-dependent proinflammatory cytokines-induced signalling pathways in the rats with abdominal artery constriction. Biomed Pharmacother 2019 Jan;109:1907-1914 (PubMed: 30551445) [IF=3.743]

9). Chen X et al. Nirogacestat suppresses RANKL-Induced osteoclast formation in vitro and attenuates LPS-Induced bone resorption in vivo. Exp Cell Res 2019 Jun 15 (PubMed: 31211955) [IF=3.329]

10). Yuan H et al. Osthole alleviated diabetic neuropathic pain mediated by the P2X4 receptor in dorsal root ganglia. Brain Res Bull 2018 Aug 14;142:289-296 (PubMed: 30118750) [IF=3.103]

11). Guo J et al. Involvement of P2Y12 receptor of stellate ganglion in diabetic cardiovascular autonomic neuropathy. Purinergic Signal 2018 Aug 6 (PubMed: 30084083) [IF=3.038]

12). Zhu L et al. miR-199b-5p Regulates Immune-Mediated Allograft Rejection after Lung Transplantation Through the GSK3β and NF-κB Pathways. Inflammation 2018 Aug;41(4):1524-1535 (PubMed: 29779167)

13). Feng Y et al. Chronic colitis induces meninges traffic of gut-derived T cells, unbalances M1 and M2 microglia/macrophage and increases ischemic brain injury in mice. Brain Res 2018 Nov 13 (PubMed: 30445026)

14). Feng Y et al. Resveratrol attenuates neuronal autophagy and inflammatory injury by inhibiting the TLR4/NF-κB signaling pathway in experimental traumatic brain injury. Int J Mol Med 2016 Apr;37(4):921-30 (PubMed: 26936125)

Application: WB    Species:rat;    Sample:Not available


15). Feng Y et al. Protective Role of Apocynin via Suppression of Neuronal Autophagy and TLR4/NF-κB Signaling Pathway in a Rat Model of Traumatic Brain Injury. Neurochem Res 2017 Nov;42(11):3296-3309 (PubMed: 28786047)

Application: WB    Species:rat;    Sample:Not available


16). Chen Y et al. NLRC4 inflammasome activation regulated by TNF-α promotes inflammatory responses in nonalcoholic fatty liver disease. Biochem Biophys Res Commun 2019 Apr 9;511(3):524-530 (PubMed: 30824190)

17). Li S et al. Early supplemental α2-macroglobulin attenuates cartilage and bone damage by inhibiting inflammation in collagen II-induced arthritis model. Int J Rheum Dis 2019 Jan 4 (PubMed: 30609267)

18). Sheng X et al. Effects of Baicalin on Diabetic Cardiac Autonomic Neuropathy Mediated by the P2Y12 Receptor in Rat Stellate Ganglia. Cell Physiol Biochem 2018;46(3):986-998 (PubMed: 29669327)

19). Zeng SY et al. Rutaecarpine prevents hypertensive cardiac hypertrophy involving the inhibition of Nox4-ROS-ADAM17 pathway. J Cell Mol Med 2019 Apr 5 (PubMed: 30953402)

20). et al. Fungal Proteins from Hericium Erinaceus Show Auxiliary Antitumor Effects with 5-Fluoro-2,4(1H,3H)-Pyrimidinedione by Improving the Gut Microbiota in Mice.

21). et al. Fungal Proteins from Hericium Erinaceus Show Auxiliary Antitumor Effects with 5-Fluoro-2, 4 (1H, 3H)-Pyrimidinedione by Improving the Gut Microbiota in Mice.

22). et al. Influences of the TLR4/NF-κB pathway on memory function and inflammatory factors in rats with cerebral small vessel disease.

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Catalog Number :

AF7014-BP
(Blocking peptide available as AF7014-BP)

Price/Size :

$200/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Pig
100%
Sheep
90%
Dog
90%
Bovine
90%
Horse
78%
Rabbit
0%
Chicken
0%
Xenopus
0%
Zebrafish
0%
High similarity Medium similarity Low similarity No similarity
P01375 as Substrate
Site PTM Type Enzyme
S2 Phosphorylation Q8N165 (PDIK1L)
K19 Myristoylation
K20 Myristoylation
S80 O-Glycosylation
S162 Phosphorylation
S171 Phosphorylation
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.