• Product name

  • Description

    Rabbit polyclonal to TNF alpha
  • Host species

  • 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.



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.


  • 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

    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 localization

    Secreted 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
    • Tnf 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 (TNF superfamily member 2) antibody
    • Tumor necrosis factor alpha antibody
    • Tumor necrosis factor antibody
    • Tumor necrosis factor ligand superfamily member 2 antibody
    • Tumor Necrosis Factor, Membrane Form antibody
    • Tumor necrosis factor, soluble form antibody
    see all


  • Anti-TNF alpha antibody (ab9635) at 0.2 µg/ml + SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate at 10 µg

    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.

  • 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.
  • 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.

  • Anti-TNF alpha antibody (ab9635) at 0.2 µg/ml + Recombinant Human TNF alpha protein (ab55237) at 0.01 µg

    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 10 seconds


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) »
See all 28 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A


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.
The following paper may of use to you for the ELISA protocol used with this antibody:

Our references for Anti-Human TNF-α (500-P31A) include one article citing use in ELISA:
Halin, C, Synergistic therapeutic effects of a tumor targeting antibody fragment, fused to interleukin 12 and to tumor necrosis factor alpha. Cancer Research; 63(12):3202-3210. 15 Jun 2003 (PubMed ID: 12810649).

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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] (https://www.abcam.com/ab8348) as capture + Rabbit polyclonal to TNF alpha (https://www.abcam.com/ab9635) as detection. Please note that these 2 antibodies are unconjugated.
Anti-TNF alpha antibody [2C8] (https://www.abcam.com/ab8348) as capture + Anti-TNF alpha antibody [F6C5 ] (HRP) (https://www.abcam.com/ab24473) as detection.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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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 detected (< 34 %) so we wouldn't anticipate cross-reactivity. Plus this is produced from sera of rabbits pre-immunized with highly pure recombinant Human TNF-α and it was purified by affinity chromatography employing immobilized Human TNF-α matrix.

We have some references for both antibodies that you may be interested in:



The protocol for using the antibodies together in sELISA can be found here: https://www.abcam.com/ps/products/8/ab8348/documents/Protocol%20rabbit.pdf

The ELISA kits we sell may come from different sources, and sometimes we are not provided with the individual antibodies for sale. We can always check individual kits to see if we have the antibodies, but in most cases they probably aren't available.

I hope this information helps. Please contact us with any other questions.

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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 recontacter si vous avez besoin de plus d'informations. Je vous souhaite bon succès dans votre recherche.

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Human Recombinant protein (expressed in E.coli strain BL21(DE3))
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

Abcam user community

Verified customer

Submitted Oct 12 2010

Western blot
Escherichia coli Purified protein (BL21(DE3))
Loading amount
1 µg
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

Abcam user community

Verified customer

Submitted Aug 26 2010

Western blot
Human Recombinant protein (Recombinant Human TFN-alpha (Source: E coil))
Loading amount
0.5 µg
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

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 overnight. 2) In separate plate, serial dilute a-h-TNF-a in assay media containing 2mg/ml Actinomycin D and 10% horse serum in triplicate. Add 60ml/well assay media containing 2ng/ml h-TNF-a resulting in 1:2 dilution. Incubate 1 hour. 3) Transfer 100ml/well h-TNF-a/anti-h-TNF-a complex to cells in assay plate Final volume/well is 200ml containing 1mg/ml Actinomycin D, 0.5 ng/ml h-TNF-a and anti-h-TNF-a as indicated below. 4) Incubate 20 hours. 5) Add 20ml of Promega Substrate CellTiter 96 Aqueous One Solution Reagent to each well. 6) Incubate 37oC and read OD at 490 nm. Data: Conc. anti-h-TNF-a O.D. reading at 490nm after 4 hours ug/ml row 1 row 2 row 3 Ave. O.D. Net O.D. 25.00 12.50 6.25 3.13 1.56 0.78 0.39 0.20 0.10 0.00 I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

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