Anti-TNF alpha antibody (ab66579)
Key features and details
- Rabbit polyclonal to TNF alpha
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-TNF alpha antibody
See all TNF alpha primary antibodies -
Description
Rabbit polyclonal to TNF alpha -
Host species
Rabbit -
Specificity
Ab66579 has been batch tested in Western Blot using LPS stimulated lysates. Abcam does not recommend this product for use in IHC. Please contact Abcam Scientific Support for more information. -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Cat, Chimpanzee, Baboon
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Immunogen
Synthetic peptide corresponding to Human TNF alpha aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab66578) -
Positive control
- This antibody gave a positive signal in the following LPS stimulated lysates: THP1, RAW 264.7 ICC/IF: PMA/Befreldin/LPS treated RAW 264.7 cell line.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab66579 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (5) |
Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 25 kDa).
Abcam recommends using 3% Milk as the blocking agent. |
| ICC/IF | (2) |
Use a concentration of 1 µg/ml.
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| Notes |
|---|
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 25 kDa). Abcam recommends using 3% Milk as the blocking agent. |
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ICC/IF
Use a concentration of 1 µg/ml. |
Target
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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. -
Involvement in disease
Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis). -
Sequence similarities
Belongs to the tumor necrosis factor family. -
Post-translational
modificationsThe soluble form derives from the membrane form by proteolytic processing.
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. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 7124 Human
- Entrez Gene: 21926 Mouse
- Entrez Gene: 24835 Rat
- Omim: 191160 Human
- SwissProt: P19101 Cat
- SwissProt: Q8HZD9 Chimpanzee
- SwissProt: P01375 Human
- SwissProt: P06804 Mouse
see all -
Alternative names
- APC1 antibody
- APC1 protein antibody
- Cachectin antibody
see all
Images
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Western blot - Anti-TNF alpha antibody (ab66579)All lanes : Anti-TNF alpha antibody (ab66579) at 1 µg/ml
Lane 1 : THP1 cells treated with PMA (negative control)
Lane 2 : THP1 cells treated with PMA+LPS+Brefeldin A
Lane 3 : Raw264.7 cells treated with PMA (negative control)
Lane 4 : Raw264.7 cells treated with PMA+LPS+Brefeldin A
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Additional bands at: 28 kDa (possible glycosylated form), 50 kDa, 75 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutesMature THP-1 and Raw264.7 cells were treated with LPS (1 ug/ml) for 6 hours to stimulate production of TNF alpha. Brefeldin A (300 ng/ml) was added in the final 3 hours of incubation to inhibit golgi traffic.
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Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody (ab66579)ab66579 staining TNF alpha in Raw264.7 cells. The cells were treated with LPS for 7 hours and Brefeldin A (ab120299) for the final 3 hours. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab66579 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (102)
ab66579 has been referenced in 102 publications.
- Li X et al. miR-142-3p targets AC9 to regulate sciatic nerve injury-induced neuropathic pain by regulating the cAMP/AMPK signalling pathway. Int J Mol Med 47:561-572 (2021). PubMed: 33416140
- Jin Q et al. Allisartan isoproxil attenuates oxidative stress and inflammation through the SIRT1/Nrf2/NF-?B signalling pathway in diabetic cardiomyopathy rats. Mol Med Rep 23:N/A (2021). PubMed: 33495841
- Parekh SV et al. Hippocampal TNF-a Signaling Mediates Heroin Withdrawal-Enhanced Fear Learning and Withdrawal-Induced Weight Loss. Mol Neurobiol 58:2963-2973 (2021). PubMed: 33580871
- Derbis M et al. Short antisense oligonucleotides alleviate the pleiotropic toxicity of RNA harboring expanded CGG repeats. Nat Commun 12:1265 (2021). PubMed: 33627639
- Zhang J et al. Minocycline attenuation of rat corpus callosum abnormality mediated by low-dose lipopolysaccharide-induced microglia activation. J Neuroinflammation 18:100 (2021). PubMed: 33902641