Anti-TNF alpha antibody (ab9635)
- Datasheet
- References (28)
- Protocols
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 -
Tested applications
Suitable for: IHC-P, WB, ELISA, Neutralising, Sandwich ELISAmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment corresponding to Human TNF alpha aa 77-233. E.coli derived Recombinant Human TNF-alpha
Database link: P01375 -
Positive control
- WB: Recombinant human TNF alpha protein (ab9642), SW480 cell lysate. IHC-P: Human tonsil tissue.
Properties
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Form
Lyophilised:Reconstitute with 200µl of sterile water. -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. -
Storage buffer
PBS, pH 7.4, no preservative, sterile filtered -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Light chain type
unknown -
Research areas
Associated products
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Compatible Secondaries
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ELISA pair antibody
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
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sELISA pair antibody
Applications
Our Abpromise guarantee covers the use of ab9635 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 |
|---|---|---|
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
| WB | Use at an assay dependent concentration. To detect TNF-alpha by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant TNF-alpha is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions. | |
| ELISA | Use at an assay dependent concentration. Can be paired for ELISA with Mouse monoclonal to TNF alpha (ab9348). To detect TNF-alpha by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant TNF-alpha. | |
| Neutralising | Use at an assay dependent concentration. To yield one-half maximal inhibition [ND50] of the biological activity of hTNF-alpha (0.5 ng/ml), a concentration of 0.08 - 0.10 µg/ml of this antibody is required. | |
| Sandwich ELISA | Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [2C8] to TNF alpha (ab8348). For sandwich ELISA, use this antibody as Detection at 0.5µg/ml with ab8348 as Capture. |
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
- Omim: 191160 Human
- SwissProt: P01375 Human
- Unigene: 241570 Human
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Alternative names
- APC1 antibody
- APC1 protein antibody
- Cachectin antibody
see all
Images
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Western blot - Anti-TNF alpha antibody (ab9635)Anti-TNF alpha antibody (ab9635) at 0.2 µg/ml + SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Observed band size: 26 kDa why is the actual band size different from the predicted?
Additional bands at: 48 kDa. We are unsure as to the identity of these extra bands. -
Sandwich ELISA - Anti-TNF alpha antibody (ab9635)Standard curve for TNF alpha (Analyte: ab9642); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [2C8] to TNF alpha (ab8348) at 5µg/ml and Detector Antibody Rabbit polyclonal to TNF alpha (ab9635) at 0.5µg/ml. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNF alpha antibody (ab9635)IHC image of TNF alpha staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9635, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Western blot - Anti-TNF alpha antibody (ab9635)
Datasheets and documents
References
This product has been referenced in:
- Yang Q et al. Adiponectin protects against uric acid-induced renal tubular epithelial inflammatory responses via the AdipoR1/AMPK signaling pathway. Int J Mol Med 43:1542-1552 (2019). Read more (PubMed: 30664190) »
- Kong YY et al. Nicotinamide phosphoribosyltransferase aggravates inflammation and promotes atherosclerosis in ApoE knockout mice. Acta Pharmacol Sin N/A:N/A (2019). Read more (PubMed: 30833708) »
