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  1. Link

    tnf-alpha-antibody-ab9635.pdf

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Immunology Innate Immunity Cytokines TNF Superfamily
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Anti-TNF alpha antibody (ab9635)

  • Datasheet
Reviews (3)Q&A (5)References (47)

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Sandwich ELISA - Anti-TNF alpha antibody (ab9635)

    Key features and details

    • Rabbit polyclonal to TNF alpha
    • Suitable for: Sandwich ELISA
    • Reacts with: Recombinant fragment
    • Isotype: IgG

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    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: Sandwich ELISAmore details
    • Species reactivity

      Reacts with: Recombinant fragment
    • Immunogen

      Recombinant fragment within Human TNF alpha aa 50 to the C-terminus. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements. E.coli derived Recombinant Human TNF-alpha
      Database link: P01375

    • 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

      Lyophilized:Reconstitute with 200µl of sterile water.
    • Storage instructions

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
    • Storage buffer

      No preservative, sterile filtered
    • Concentration information loading...
    • Purity

      Immunogen affinity purified
    • Clonality

      Polyclonal
    • Isotype

      IgG
    • Light chain type

      unknown
    • 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

    • ELISA pair antibody

      • Anti-TNF alpha antibody [2.1_4E10-1H11] (ab9348)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
    • Positive Controls

      • Recombinant human TNF alpha protein (ab9642)
    • Recombinant Protein

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

      • Human TNF alpha ELISA Kit (ab181421)
      • Human TNF alpha Matched Antibody Pair Kit (ab213467)

    Applications

    The Abpromise guarantee

    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
    Sandwich ELISA
    Use a concentration of 0.5 - 2 µg/ml.

    To detect Human TNF-α by sandwich ELISA (using 100 μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of this antibody is required.

    Notes
    Sandwich ELISA
    Use a concentration of 0.5 - 2 µg/ml.

    To detect Human TNF-α by sandwich ELISA (using 100 μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of this antibody is required.

    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
    • 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

    • Sandwich ELISA - Anti-TNF alpha antibody (ab9635)
      Sandwich ELISA - Anti-TNF alpha antibody (ab9635)

      ab9635 detecting TNF alpha with sandwich ELISA

    Protocols

    • Western blot protocols
    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (47)

    Publishing research using ab9635? Please let us know so that we can cite the reference in this datasheet.

    ab9635 has been referenced in 47 publications.

    • Zhu J  et al. MicroRNA-140-5p regulates the proliferation, apoptosis and inflammation of RA FLSs by repressing STAT3. Exp Ther Med 21:171 (2021). PubMed: 33456538
    • Mohamed IN  et al. Deletion of Thioredoxin-Interacting Protein (TXNIP) Abrogates High Fat Diet-induced Retinal Leukostasis, Barrier Dysfunction and Microvascular Degeneration in a Mouse Obesity Model. Int J Mol Sci 21:N/A (2020). PubMed: 32492941
    • Korgaonkar AA  et al. Distinct cellular mediators drive the Janus faces of toll-like receptor 4 regulation of network excitability which impacts working memory performance after brain injury. Brain Behav Immun 88:381-395 (2020). PubMed: 32259563
    • Menga A  et al. Glufosinate constrains synchronous and metachronous metastasis by promoting anti-tumor macrophages. EMBO Mol Med N/A:e11210 (2020). PubMed: 32885605
    • Liu T  et al. LncRNA THRIL is upregulated in sepsis and sponges miR-19a to upregulate TNF-a in human bronchial epithelial cells. J Inflamm (Lond) 17:31 (2020). PubMed: 32944003
    View all Publications for this product

    Customer reviews and Q&As

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

    1-8 of 8 Abreviews or Q&A

    ELISA abreview for Anti-TNF alpha antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Recombinant protein (expressed in E.coli strain BL21(DE3))
    Specification
    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
    Type
    Indirect
    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 Oct 12 2010

    Western blot abreview for Anti-TNF alpha antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Escherichia coli Purified protein (BL21(DE3))
    Loading amount
    1 µg
    Specification
    BL21(DE3)
    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
    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 Aug 26 2010

    Western blot abreview for Anti-TNF alpha antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Recombinant protein (Recombinant Human TFN-alpha (Source: E coil))
    Loading amount
    0.5 µg
    Specification
    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
    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 Feb 15 2010

    Question

    Kindly contacted us for informaton about use of these products in ELISA.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 13 2013

    Answer

    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).

    Read More

    Kevin Hanson

    Abcam Scientific Support

    Answered on Sep 13 2013

    Question

    Inquiry: Hello We would like to develop an Electrochemical Immunosensor using a sandwich type assay (Primary Antibody-Antigen- Secondary antibody) and we need to choose a suitable pair of Ab with high affinity reaction with their Antigen: TNF alpha protein for Immunoassay applications.. We would like to know the code of suitable primary and secondary antibodies. that you recommend us Thank you very much. Regards

    Read More

    Abcam community

    Verified customer

    Asked on Oct 22 2012

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 22 2012

    Question

    On your website you give AB9635 as a good candidate for sandwich assay with AB8348. I would like to use that combination in my PhD. The goal is to detect TNF-alpha in cell culture medium with high specificity and even when the antigen is present in low concentrations. That’s why information about cross reactivity is important. Do you have any information about proteins or other molecules that can cross react with the antibody pair?

    I was also wondering if there is a reason that the ELISA-kits don’t use the antibodies you sell individually. It would be a nice comparison to make use of an ELISA kit with the AB8348 and AB9635 antibodies.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 12 2012

    Answer

    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:

    https://www.abcam.com/TNF-alpha-antibody-ab9635-references.html

    https://www.abcam.com/TNF-alpha-antibody-2C8-ab8348-references.html

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 12 2012

    Question

    searching for antibody blocking TNF alpha on human cultured cells - blocking free and membrane bound?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 04 2011

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 04 2011

    Question

    Can you tell me how the neutralization experiment was done?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 15 2005

    Answer

    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,

    Read More

    Abcam Scientific Support

    Answered on Jul 22 2005

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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