• Product nameAnti-TNF alpha antibody
    See all TNF alpha primary antibodies
  • Description
    Rabbit polyclonal to TNF alpha
  • SpecificityAb66579 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 applicationsSuitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cat, Chimpanzee, Baboon
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human TNF alpha.

    (Peptide available as ab66578.)

  • Positive control
    • This antibody gave a positive signal in the following LPS stimulated lysates: THP1, Raw264.7 IF/ICC: PMA/Befreldin/LPS treated RAW246.7 cell line.



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 Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 25 kDa).Can be blocked with Human TNF alpha peptide (ab66578).

Abcam recommends using 3% Milk as the blocking agent.

ICC/IF Use a concentration of 1 µg/ml.


  • 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
    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
    • Tnf 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, Membrane Form antibody
    • Tumor necrosis factor, soluble form antibody
    see all

Anti-TNF alpha antibody images

  • 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.

    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 minutes

    Mature 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.

  • ab66579 stained RAW246.7 cells. The cells were treated with PMA for 24 hours, LPS for 6 hours and Befreldin for the final 3 hours. The cells were then 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66579 at 1µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

References for Anti-TNF alpha antibody (ab66579)

This product has been referenced in:
  • Riemann A  et al. Acidic environment activates inflammatory programs in fibroblasts via a cAMP-MAPK pathway. Biochim Biophys Acta 1853:299-307 (2015). Read more (PubMed: 25461841) »
  • Ilievski V  et al. TLR4 Expression by Liver Resident Cells Mediates the Development of Glucose Intolerance and Insulin Resistance in Experimental Periodontitis. PLoS One 10:e0136502 (2015). IF ; Mouse . Read more (PubMed: 26317345) »

See all 11 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (Placenta)
Gel Running Conditions Reduced Denaturing
Loading amount 20 µg
Specification Placenta
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C


Verified customer

Submitted Dec 22 2016

Application Western blot
Sample Rat Tissue lysate - whole (LIVER)
Gel Running Conditions Reduced Denaturing
Loading amount 50 µg
Specification LIVER
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 06 2016

Thank you for contacting us.

The antibodies ab66579 and ab71281 are polyclonal antibodies and are guaranteed form detection of mouse TNF alpha protein. We do not determine the affinity of polyclonal antibodies which is rather difficult because...

Read More

Thank you for your enquiry.

I am sorry to confirm that most of our TNF alpha antibodies, including ab17611, have an epitope within the soluble region of the protein.

I have found one antibody, tested in mouse, that has an immunoge...

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Please check the online datasheets of all antibodies. You will find the required information.


Thank you very much for contacting us.

The possible rabbit specific antibodies are available as;

ICAM1: the rabbit protein sequence (http://www.uniprot.org/uniprot/Q75T02) does not show any homology with human protein (http:...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Liver)
Loading amount 20 µg
Specification Liver
Gel Running Conditions Non-reduced Denaturing (15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 10 2012

Vielen Dank für Ihren Anruf.

Wie am Telefon besprochen, habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 1234567 und solltespätestens Übermorgenbei Ihnen ankommen.

Read More

Thank you for contacting us.

The IHC-Fr data was provided by a customer in an Abreview. You can see that here:


We do not know of any pu...

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I am glad you followed up with me as I have searched our records and unfortunately we did not receive the email you sent on 12/5/2011.

I also apologize that my reply was not helpful for you. I would be happy to hear of any suggestions to help...

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1-10 of 16 Abreviews or Q&A