Human TNF alpha ELISPOT Set (ab46550)

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Key features and details

  • Assay type: Sandwich (qualitative)
  • Sample type: Suspension cells
  • Reacts with: Human, Macaque monkey

Overview

  • Product name

    Human TNF alpha ELISPOT Set
    See all TNF alpha kits

  • Sample type

    Suspension cells

  • Assay type

    Sandwich (qualitative)

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Human, Macaque monkey

  • Product overview

    Abcam’s Human TNF alpha ELISPOT Set is an in vitro ELISPOT assay designed for the qualitative measurement of TNFα production and secretion in a single cell suspension.


     


    The ELISPOT assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. ELISPOT assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.



    The ELISPOT assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.


     


    After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.


     


    Recognizes natural human TNF alpha.

  • Tested applications

    Suitable for: ELISpotmore details

  • Platform

    Reagents

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 5 x 96 tests 10 x 96 tests 15 x 96 tests 20 x 96 tests
    Bovine Serum Albumin 1 x 1g 2 x 1g 3 x 1g 4 x 1g
    Ready-to-use BCIP/NBT substrate buffer 1 x 50ml 2 x 50ml 3 x 50ml 4 x 50ml
    Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl 2 x 50µl 3 x 50µl 4 x 50µl
    TNFα Biotinylated detection antibody (clone B-C7) 1 vial 2 vials 3 vials 4 vials
    TNFα Capture antibody (clone B-F7) 1 x 0.5ml 2 x 0.5ml 3 x 0.5ml 4 x 0.5ml

  • Research areas

  • 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

  • Alternative names

    • APC1
    • APC1 protein
    • Cachectin
    • DIF
    • Differentiation inducing factor
    • Macrophage cytotoxic factor
    • Tnf
    • TNF superfamily member 2
    • TNF superfamily, member 2
    • TNF, macrophage derived
    • TNF, monocyte derived
    • TNF-a
    • TNF-alpha
    • TNFA
    • TNFA_HUMAN
    • TNFSF2
    • Tumor necrosis factor
    • Tumor necrosis factor (TNF superfamily member 2)
    • Tumor necrosis factor alpha
    • Tumor necrosis factor ligand superfamily member 2
    • Tumor Necrosis Factor, Membrane Form
    • Tumor necrosis factor, soluble form
    see all

  • Database links

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab46550 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
    ELISpot
    Use at an assay dependent dilution.
    Notes
    ELISpot
    Use at an assay dependent dilution.

    References (1)

    Publishing research using ab46550? Please let us know so that we can cite the reference in this datasheet.

    ab46550 has been referenced in 1 publication.

    • Dissanayake K  et al. Potential applicability of cytokines as biomarkers of disease activity in rheumatoid arthritis: Enzyme-linked immunosorbent spot assay-based evaluation of TNF-a, IL-1ß, IL-10 and IL-17A. PLoS One 16:e0246111 (2021). PubMed: 33497394

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