Overview

  • Product name
  • Description
    Rabbit polyclonal to TNF alpha
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, WB, IHC-P, ELISA, RIA, Neutralisingmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Recombinant full length protein corresponding to Mouse TNF alpha.
    (Peptide available as ab151311)

Properties

Applications

Our Abpromise guarantee covers the use of ab34674 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
ICC/IF 1/200.
IP 1/400 - 1/800.
WB 1/400 - 1/800. Detects a band of approximately 17 kDa (predicted molecular weight: 26 kDa).Can be blocked with Synthetic Mouse TNF alpha protein (ab151311). Detects TNF alpha from supernatants or lysates of 2 x 106 endotoxin stimulated mouse peripheral blood mononuclear cells (PBMC).
IHC-P Use at an assay dependent concentration.
ELISA 1/1000 - 1/2000. For use in ELISA formats, this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody.
RIA 1/8000.
Neutralising 1/200. It is recommended to incubate the sample with the dilution of the antibody for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG is recommended.

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

  • Anti-TNF alpha antibody (ab34674) at 1/1000 dilution
    Secondary
    HRP rabbit secondary antibody at 1/40000 dilution

    Predicted band size: 26 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?

  • ab34674 staining TNF alpha in immortalised bone marrow-derived macrophages by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde and permeabilized in 0.1% Triton X-100 prior to blocking in 1% BSA + 5% goat serum in PBS for 1 hour at 21°C. The primary antibody was diluted 1/200 in the blocking buffer and incubated with the sample for 1 hour at 21°C. The secondary antibody was Alexa Fluor® 568-conjugated Goat anti-Rabbit polyclonal, diluted 1/500.

    See Abreview

  • ab34674 staining TNF alpha in mouse knee joint sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 10 minutes at room temperature followed by incubation with the primary antibody at a 1/200 dilution for 30 minutes. An undiluted HRP-conjugated human anti-rabbit monoclonal was used as secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Qu W  et al. Inhibition of colony-stimulating factor 1 receptor early in disease ameliorates motor deficits in SCA1 mice. J Neuroinflammation 14:107 (2017). Read more (PubMed: 28545543) »
  • Rozas P  et al. Targeted overexpression of tumor necrosis factor-a increases cyclin-dependent kinase 5 activity and TRPV1-dependent Ca2+ influx in trigeminal neurons. Pain 157:1346-62 (2016). Read more (PubMed: 26894912) »
See all 15 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Rabbit Tissue lysate - whole (aortic tissue)
Specification
aortic tissue
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Skin)
Permeabilization
Yes - Triton X-100
Specification
Skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 01 2013

Question
Answer

Vielen Dank für Ihre Rückmeldung.

Das Produkt ist unter "TNF alpha peptide (ab151311) " zu finden.

https://www.abcam.com/tnf-alpha-peptide-ab151311.html

Ich hoffe der Link hilft Ihnen weiter. Sollten Sie weiterhin Probleme haben das Produkt zu finden zögern sie bitte nicht sich wieder bei mir zu melden.

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Answer

Ich freue mich Ihnen mitteilen zu können, dass das TNF alpha peptide (ab151311) jetzt in unseren Produktkatalog aufgenommen wurde.Sie können das Produkt wie gewohnt bei uns Bestellen.

Ich hoffe ich konnte Ihnen helfen. Wenn Sie weitere Fragen haben können Sie gerne wieder auf mich zurückkommen.

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Answer

Vielen Dank für Ihre email.

Ich bin mit dem Labor in Kontakt und hoffe das wir Ihnen das zum Anti-TNF alpha antibody (ab34674)gehörende Peptide bald zum Verkauf anbieten können.

Sobald ich die Bestätigung aus dem Labor dafür erhalte werde ich mich bei Ihnen melden.

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Answer

Vielen Dank für Ihre Anfrage.

Ich habe mich wegen des TNF alpha Peptides an das Labor gewendet und werde sie informieren sobald ich eine Rückmeldung bekomme ob wir dieses zum Verkauf anbieten können.

Wir legen sehr viel Wert auf unsere Qualitätskontrolle. Von daher wäre es hilfreich, wenn sie einen Fragebogen zu dem Problem für uns ausfüllen könnten. Mit Hilfe des Fragebogens können wir Daten zu unseren Produkten sammeln und so Probleme mit unseren Produkten frühzeitig erkennen und analysieren.

Order Details
Antibody code:

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)


Optimization attempts (problem solving)
How many times have you tried the IHC?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


Es wäre außerdem hilfreich wenn sie uns ein Bild von Ihrem Ergebnis zusenden könnten.

Vielen herzlichen Dank für Ihre Zeit. Ich freue mich auf Ihre Antwort.

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Answer

Thank you for your email.

There may be some antibodies where I might have suggested the human tissues or cells. I apologize for this. I have included the positive control information irrespective of species so similar mouse tissues sections can used as positive control.

Actually the positive control information for each antibody is present on the datasheet; please check the datasheet. Alternatively, we always follow human protein Atlas database (http://www.proteinatlas.org/index.php). The second option is, searching positive control for other antibodies in catalogue against same target; the cell line or tissue sections tested with those antibodies could also be used as positive control. The third place to search for is literature.

I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

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Answer

Thank you for contacting us.

The positive control for ab36392 is IL2 activatedcultured NK cells.

ab9918; human tonsil sections, human cervical squamous cell carcinoma section

ab34674; human artery or colon tissue sections

ab15537; HepG2 cells. mouse brain sections, human placenta sections; Human hepatocellular liver carcinoma cell line; Human hepatocarcinoma tissue sections

ab2557; mouse spleen tissue sections

ab9475; human tonsil tissue sections

ab22378; mouse spleen tissue sections

ab51312; Thymus tissue sections, human pancreas tissue sections.

The best database to search for positive control is Human Protein Atlas. Please click the link http://www.proteinatlas.org/index.php

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

hiermit sende ich ihnen den ausgefüllten Fragebogeb zurück, und hoffe sie können mir mit ihrer Antwort weiterhelfen.
Vielen Dank


1) Abcam product code ab
TGF ß 66043
TNF alpha 34674
IL-6 6672

2) Abcam order reference number or product batch number

2) Description of the problem
Habe die Kerne DAPI gefärbt in Kombination mit TGF ß, TNF alpha oder IL-6,
Kernfärbung war zu sehen, aber nicht die anderen AK, obwohl das gefärbte Gewebe TGF ß, TNF alpha und IL-6 expremiert war

4) Sample preparation:
Species mouse
Type of sample:/formalin fixed paraffin embedded sections
Sample preparation
Positive control
Negative control

5) Fixation step
Yes
If yes: Fixative agent and concentration Formalin 4%
Fixation time über Nacht
Fixation temperature Raumtemperatur

6) Antigen retrieval method
Citratpuffer pH 6.0

7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
Permeabilizing agent and concentration: Citratpuffer enthält Tween 20


8) Blocking agent (eg BSA, serum…): 1% BSA, 2% goat serum
Concentration
Blocking time 20 min
Blocking temperature Raumtemperatur

9) Endogenous peroxidases blocked?
Endogenous biotins blocked?

10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution TGF ß :1,5,10 µg/ml TNF alpha: 2.2, 11, und 22µg/ml, IL-6: 160, 800 und 1600µg/ml
Diluent buffer : Blockingpuffer
Incubation time Über Nacht 4°C

11) Secondary antibody:
Species:goat anti rabbit IgG
Reacts against:
Concentration or dilution 1:500
Diluent buffer 0,1% Triton X100, 0,05% Tween 20 in PBS
Incubation time 1h
Fluorochrome or enzyme conjugate 488

12) Washing after primary and secondary antibodies:
Buffer
Number of washes 3x

13) Detection method

14) How many times have you run this staining?
Do you obtain the same results every time?
What steps have you altered to try and optimize the use of this antibody?

Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.

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Answer

Vielen Dank für Ihre Antwortund dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

Es tut mir leid, dass Sie Probleme mit unseren Antikörpern hatten.

Um Ihr Problem zu lösen, möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen:

Es ist sehr unwahrscheinlich, dass drei Antikörper gegen verschieden Zielprotein nicht funktionieren. Daher bin ich zuversichtlich, dass mit Protokolloptimierung eine Besserung der Ergebnisse erreicht werden kann.

1.) Der gleiche sekundäre Antikörper wird fier alle drei primären Antikörper benutzt. Funktioniert dieser mit anderen primären? Ich schlage vor einen anderen sekundären auszuprobieren, am besten auch mit einem anderen Fluorochrom um Probleme mit dem Mikroskop auszuschließen.

2.) Für die meisten Zielproteine ist Citrate Puffer als Antigendemaskierung suboptimal. ich schlage deshalb vor EDTA, pH 9 auszuprobieren. ich habe mehr Infomaterial als Attachement angehängt.

3.) Da es sich um sekretierte Proteine handelt ist eine Permeabilisierung nicht unbedingt nötig. Ich schlage vor allem vor, nicht Triton und Tween in der Lösung für den sekundären Antikörper zu verwenden. So kann TBS 0.025% Triton als Puffer für die Antikörper verwendet werden.

4.) Bei zu wenig oder fehlendem Signal können die primären Antikörper auch nur in TBS 0.025% Triton inkubiert werden. Ich stimme ihnen zu, dass über Nacht bei 4C die beste Option ist.

Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte und zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab34674. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

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1-10 of 15 Abreviews or Q&A

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