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    products/primary-antibodies/tnf-alpha-antibody-ab34674.pdf

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Immunology Innate Immunity Cytokines TNF Superfamily
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Anti-TNF alpha antibody (ab34674)

  • Datasheet
Reviews (5)Q&A (10)References (40)

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Abpromise

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Find out more.

Western blot - Anti-TNF alpha antibody (ab34674)

    Key features and details

    • Rabbit polyclonal to TNF alpha
    • Suitable for: IP, RIA, ICC/IF, WB, Neutralising, IHC-P, ELISA
    • Reacts with: Mouse, Recombinant fragment
    • Isotype: IgG

    Get better batch-to-batch reproducibility with a recombinant antibody

    Product image
    Anti-TNF alpha antibody [EPR21753-109] (ab205587)
    • Research with confidence – consistent and reproducible results with every batch
    • Long-term and scalable supply – powered by recombinant technology for fast production
    • Success from the first experiment – confirmed specificity through extensive validation
    • Ethical standards compliant – production is animal-free

    Overview

    • Product name

      Anti-TNF alpha antibody
      See all TNF alpha primary antibodies
    • Description

      Rabbit polyclonal to TNF alpha
    • Host species

      Rabbit
    • Tested applications

      Suitable for: IP, RIA, ICC/IF, WB, Neutralising, IHC-P, ELISAmore details
    • Species reactivity

      Reacts with: Mouse, Recombinant fragment
    • Immunogen

      Recombinant full length protein corresponding to Mouse TNF alpha.

    • General notes

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
    • Storage buffer

      Preservative: 0.01% Sodium azide
      Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
    • Concentration information loading...
    • Purity

      Whole antiserum
    • Clonality

      Polyclonal
    • Isotype

      IgG
    • Research areas

      • Immunology
      • Innate Immunity
      • Cytokines
      • TNF Superfamily
      • Signal Transduction
      • Growth Factors/Hormones
      • TNF
      • Microbiology
      • Organism
      • Virus
      • RNA Virus
      • ssRNA positive strand virus
      • SARS Coronavirus
      • Cancer
      • Growth factors
      • TNF
      • Cardiovascular
      • Atherosclerosis
      • Vascular Inflammation
      • Inflammatory mediators
      • Metabolism
      • Types of disease
      • Metabolic disorders
      • Neuroscience
      • Development
      • Neuroscience
      • Diseases
      • Neuroscience
      • Processes

    Associated products

    • Conjugation kits

      • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
      • APC Conjugation Kit - Lightning-Link® (ab201807)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
    • Recombinant Protein

      • Recombinant human TNF alpha protein (Active) (ab259410)
      • Recombinant mouse TNF alpha protein (Active) (ab259411)
    • Related Products

      • Mouse TNF alpha ELISA Kit (ab208348)

    Applications

    The Abpromise guarantee

    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
    IP
    Use at an assay dependent concentration.
    RIA
    Use at an assay dependent concentration.
    ICC/IF (1)
    Use at an assay dependent concentration.
    WB (1)
    1/500 - 1/2000. Detects a band of approximately 17 kDa (predicted molecular weight: 26 kDa).
    Neutralising
    Use at an assay dependent concentration.

    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.

    IHC-P (2)
    Use at an assay dependent concentration.
    ELISA
    Use at an assay dependent concentration.

    For use in ELISA formats, this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody.

    Notes
    IP
    Use at an assay dependent concentration.
    RIA
    Use at an assay dependent concentration.
    ICC/IF
    Use at an assay dependent concentration.
    WB
    1/500 - 1/2000. Detects a band of approximately 17 kDa (predicted molecular weight: 26 kDa).
    Neutralising
    Use at an assay dependent concentration.

    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.

    IHC-P
    Use at an assay dependent concentration.
    ELISA
    Use at an assay dependent concentration.

    For use in ELISA formats, this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody.

    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.
    • Target information above from: UniProt accession P01375 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 21926 Mouse
      • SwissProt: P06804 Mouse
      • Unigene: 1293 Mouse
      • 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

      Images

      • Western blot - Anti-TNF alpha antibody (ab34674)
        Western blot - Anti-TNF alpha antibody (ab34674)
        Anti-TNF alpha antibody (ab34674) at 1/1000 dilution + recombinant mouse TNF alpha, 50ng

        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?

      Protocols

      • Immunoprecipitation protocols
      • Immunocytochemistry & immunofluorescence protocols
      • Western blot protocols

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet download

        Download

      References (40)

      Publishing research using ab34674? Please let us know so that we can cite the reference in this datasheet.

      ab34674 has been referenced in 40 publications.

      • Ertle CM  et al. New Pathways for the Skin's Stress Response: The Cholinergic Neuropeptide SLURP-1 Can Activate Mast Cells and Alter Cytokine Production in Mice. Front Immunol 12:631881 (2021). PubMed: 33815383
      • Abriz AE  et al. Effect of docosahexaenoic acid plus insulin on atherosclerotic human endothelial cells. J Inflamm (Lond) 18:10 (2021). PubMed: 33602249
      • Liu H  et al. Catalytically potent and selective clusterzymes for modulation of neuroinflammation through single-atom substitutions. Nat Commun 12:114 (2021). PubMed: 33414464
      • Sun W  et al. Tetrahydropyrimidines, ZL-5015 Alleviated Lipopolysaccharide (LPS)-Induced Acute Pneumonia in Rats by Activating the NRF-2/HO-1 Pathway. Med Sci Monit 26:e924482 (2020). PubMed: 32844782
      • Sáenz-Medina J  et al. Nox1-derived oxidative stress as a common pathogenic link between obesity and hyperoxaluria-related kidney injury. Urolithiasis 48:481-492 (2020). PubMed: 31690958
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-10 of 15 Abreviews or Q&A

      Western blot abreview for Anti-TNF alpha antibody

      Good
      Abreviews
      Abreviews
      abreview image
      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
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Jul 09 2013

      Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-TNF alpha antibody

      Excellent
      Abreviews
      Abreviews
      abreview image
      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
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Jul 01 2013

      Question

      Guten Tag,
      leider fibde ich dieses Produkt nicht auf ihrer Website.

      Read More

      Abcam community

      Verified customer

      Asked on Nov 14 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Nov 14 2012

      Question

      Guten Tag,

      grundsätzlich gibt es kein Problem, jedoch wäre das Peptid interessant um so die Spezifität des Antikörpers zu validieren. Mögliche Publikationen bedarfen einer validen negativ Kontrolle und einem Beweis über die Funktionalität des Antikörpers.

      Read More

      Abcam community

      Verified customer

      Asked on Nov 14 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Nov 14 2012

      Question

      Guten Tag,

      grundsätzlich gibt es kein Problem, jedoch wäre das Peptid interessant um so die Spezifität des Antikörpers zu validieren. Mögliche Publikationen bedarfen einer validen negativ Kontrolle und einem Beweis über die Funktionalität des Antikörpers.

      Read More

      Abcam community

      Verified customer

      Asked on Nov 13 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Nov 13 2012

      Question

      Guten Tag,
      wir nutzen den o.g. Antikörper von Ihnen und haben eine positive Färbung in Nervenfasern beobachten können. Da uns dieses Ergebnis etwas überrascht würden wir gern die Spezifität des AK mittels des passenden Peptides validieren? Gibt es bei Ihnen das passende Peptid zu diesem AK?

      Read More

      Abcam community

      Verified customer

      Asked on Nov 12 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Nov 12 2012

      Question

      One doubt: some of this antibody only react with mouse, and you suggested human tissue. Is it correct?
      Thanks,

      Read More

      Abcam community

      Verified customer

      Asked on Aug 14 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Aug 15 2012

      Question

      Our customer would like to know which are the best positive control
      and negative control for the antibodies below.
      1. anti Natural killer - ab36392
      2. anti- interferon gama - ab9918
      3. anti TNF alfa - ab34674
      4. anti TGF beta - ab15537
      5. anti neutrófilo - ab2557
      6. anti CD20 - ab9475
      7. anti células T CD8 - ab22378
      8. anti células T CD4 - ab51312
      She will do ICC and IHC-P assays.
      Thanks and best regards,

      Read More

      Abcam community

      Verified customer

      Asked on Aug 10 2012

      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

      Read More

      Abcam Scientific Support

      Answered on Aug 10 2012

      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.

      Read More

      Abcam community

      Verified customer

      Asked on Apr 24 2012

      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.

      Read More

      Abcam Scientific Support

      Answered on Apr 24 2012

      Question

      I have purchased ab34674. I have used it successfully several times in western blot to detect protein from cell supernatant. I have stored the antibody in aliquots at -20 deg C. But then I tried the experiment again and the newly thawed aliquot of antibody didn't detect any bands. all other aspects of the experiment were the same as before. this is all within a few weeks of receiving the antibody. an Actin antibody works with the lysate and the secondary and ECL is fine.

      Read More

      Abcam community

      Verified customer

      Asked on Dec 13 2011

      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.  

      Read More

      Abcam Scientific Support

      Answered on Dec 13 2011

      1-10 of 15 Abreviews or Q&A

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