Overview

  • Product name
  • Description
    Rabbit polyclonal to TNF alpha
  • 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
  • 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.

Properties

Applications

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.

Target

  • 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
    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 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, 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

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.

    Secondary
    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

This product has been referenced in:
  • Murphy RA  et al. Establishment of HK-2 Cells as a Relevant Model to Study Tenofovir-Induced Cytotoxicity. Int J Mol Sci 18:N/A (2017). Read more (PubMed: 28257038) »
  • McGavigan AK  et al. Vertical sleeve gastrectomy reduces blood pressure and hypothalamic endoplasmic reticulum stress in mice. Dis Model Mech 10:235-243 (2017). WB ; Mouse . Read more (PubMed: 28093508) »

See all 34 Publications for this product

Customer reviews and Q&As

Application
Flow Cytometry
Sample
Rat Cell (Brain)
Permeabilization
Yes - 0.25% Triton X-100
Gating Strategy
a single cells population
Specification
Brain
Fixation
Paraformaldehyde
Username

Ms. Aneta Radziwon-Balicka

Verified customer

Submitted Sep 13 2017

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
Username

Vicky

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
Username

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.

Thanks!

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
Username

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.
...

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Thank you for contacting us.

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

http://www.abcam.com/index.html?datasheet=66579&tab=abreviews&intabreviewid=23420

We do not know of any pu...

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1-10 of 17 Abreviews or Q&A

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

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