Overview

  • Product nameAnti-TNF alpha antibody
    See all TNF alpha primary antibodies
  • Description
    Rabbit polyclonal to TNF alpha
  • Tested applicationsIHC-P, WB, ELISA, Neutralising, Sandwich ELISAmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Highly pure (>98%) recombinant hTNF-alpha (human Tumor Necrosis Factor-alpha); catalogue number ab9642.

  • Positive control
    • IHC-P: FFPE human tonsil tissue sections.

Properties

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

  • FunctionCytokine 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 diseaseGenetic 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 similaritiesBelongs to the tumor necrosis factor family.
  • Post-translational
    modifications
    The 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 localizationSecreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • APC1 antibody
    • APC1 protein antibody
    • Cachectin antibody
    • DIF antibody
    • Differentiation inducing factor antibody
    • Macrophage cytotoxic factor antibody
    • MCF antibody
    • Necrosin antibody
    • TNF a antibody
    • TNF a antibody
    • TNF alpha antibody
    • TNF antibody
    • TNF antibody
    • TNF Macrophage Derived antibody
    • TNF Monocyte Derived antibody
    • TNF Superfamily Member 2 antibody
    • TNF superfamily, member 2 antibody
    • TNF, macrophage derived antibody
    • TNF, monocyte derived antibody
    • TNF-a antibody
    • TNF-alpha antibody
    • TNFA antibody
    • TNFA_HUMAN antibody
    • TNFSF2 antibody
    • Tumor necrosis factor alpha antibody
    • Tumor necrosis factor antibody
    • Tumor necrosis factor ligand superfamily member 2 antibody
    • Tumor necrosis factor ligand superfamily member 2 antibody
    • Tumor Necrosis Factor Precursor antibody
    • Tumor Necrosis Factor, Membrane Form antibody
    • Tumor necrosis factor, soluble form antibody
    • Tumour Necrosis Factor Alpha antibody
    see all

Anti-TNF alpha antibody images

  • 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.

  • 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.
  • 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.

Protocols

References for Anti-TNF alpha antibody (ab9635)

This product has been referenced in:
  • Li S  et al. Cathepsin B Contributes to Autophagy-related 7 (Atg7)-induced Nod-like Receptor 3 (NLRP3)-dependent Proinflammatory Response and Aggravates Lipotoxicity in Rat Insulinoma Cell Line. J Biol Chem 288:30094-104 (2013). WB ; Rat . Read more (PubMed: 23986436) »
  • Theilacker N  et al. Multiplexed protein analysis using encoded antibody-conjugated microbeads. J R Soc Interface : (2011). Read more (PubMed: 21247950) »

See all 2 Publications for this product

Product Wall

In order to detect Human TNF-α by sandwich ELISA (using 100μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of this antibody is required. We recommend determining optimal dilutions of this antibody for your ELISA assay.
Th...

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Thank you for contacting us.

We have in our catalogue the following pairs of antibodies that have been tested for the detection of Human TNF-alpha in sELISA :

Anti-TNF alpha antibody [2C8] (http://www.abcam.com/ab8348) as capture + Ra...

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Thank you for your inquiry.

We haven't specifically tested ab8348 and ab9635 for cross-reactivity with other proteins.

For ab9635, I BLASTed the immunogen sequence against other human proteins and no significant homology was detecte...

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Merci pour votre patience. Veuillez trouver en attaché le document que le laboratoire m'avait envoyé pour le ab9635. Il montre que l'anticorps est efficace à inhiber le TNF alpha à moins que 0.1ug/ml. Veuillez ne pas hésiter à nous reco...

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Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application ELISA
Sample Human Recombinant protein (expressed in E.coli strain BL21(DE3))
Specification expressed in E.coli strain BL21(DE3)
Blocking step Milk as blocking agent for 20 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Type Indirect
Username

Abcam user community

Verified customer

Submitted Oct 12 2010

Application Western blot
Sample Escherichia coli Purified protein (BL21(DE3))
Loading amount 1 µg
Specification BL21(DE3)
Gel Running Conditions Reduced Denaturing (4-20%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Aug 26 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Recombinant protein (Recombinant Human TFN-alpha (Source: E coil))
Loading amount 0.5 µg
Specification Recombinant Human TFN-alpha (Source: E coil)
Gel Running Conditions Reduced Denaturing (4-20%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 15 2010

Thank you for your enquiry and your patience. The neutralization experiment was conducted as follows: Procedure: 1) Resuspend 929 cells in regular culture media. Transfer 100ml/well in assay plate (8,100 cells/well). Incubate 37oC over...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"