Overview

  • Product name

    Human TNF alpha ELISA Set (without plates)
    See all TNF alpha kits
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 10 pg/ml
  • Range

    25 pg/ml - 800 pg/ml
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s TNFα Human in vitro ELISA Set is designed for the quantitative measurement of TNFα in cell culture supernatants, buffered solutions, serum, plasma and other body fluids.

     

    The TNF alpha Elipair includes both the capture and the detection antibodies as well as Horse-Radish peroxidase conjugated Streptavidin (HRP-Strep) and TMB which are the reagents used in the colour development step.

     

    This Elipair set is intended to measure human TNF alpha in cell -culture supernatants, as well as in serum and plasma samples. The enclosed procedures provide general guidelines for ELISA preparation. This procedure may need some optimization depending on the plastic, matrix, standard range.

     

    Recognizes both natural and recombinant human TNF alpha.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Reagents

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 5 x 96 tests 10 x 96 tests
    Detection Biotinylated anti-TNF alpha Antibody 1 x 0.55ml 2 x 0.55ml
    Chromogen TMB Substrate Solution 2 vials 4 vials
    Streptavidin-HRP 1 vial 2 vials
    TNF alpha Capture Antibody 1 x 0.5ml 2 x 0.5ml
    TNF alpha Standard 5 vials 10 vials
  • 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

Our Abpromise guarantee covers the use of ab47347 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 at an assay dependent concentration.

Images

  • TNF alpha Standard curve ranging from 25 to 800 pg/mL

References

ab47347 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Question
Answer


The non-biotinylated version of the detection antibody is available on our catalog as ab27381.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

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Question
Answer

The detection antibody is available as ab27381.

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Question
Answer

Thank you for your inquiry.
HRP-Streptavidin is provided at 1.0mg/ml for a final working concentration of 0.15µg/ml when diluted according to the recommendations.
I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for contacting us.

We do not sell individual components for ab47347 at the moment.

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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Answer

Thank you very much for your reply.

I am glad that you have found some kits that will be suitable for your assay. If you need any help standardizing your methods, please let me know and I'll be happy to assist.

Have a wonderful day.

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Answer

Thank you very much for contacting us with your questions, and for your patience while I have been in touch with the lab regarding your enquiry.

For the TAC kit ab65329, you can use a general lysis buffer such as RIPA to prepare the lysates.To prepare the tissue lysates:

Start with 10-100 mg of the tissue and add 500-1000 μl (or ˜4-6 volumes) of the lysis buffer on ice. Homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.

The absorbance will be measured at 570 nm and there will be a pink color development. The DMSOused to prepare the standard is not supplied with the kit.

For the TNFa kits, the Elipair product ab47347 only contains the capture and detection antibodies, the TMB substrate, streptavidin-HRP, and the TNFa standard. Using this product, you would need to set up the other components of the kit. For example, you will need to obtain 96-well plates and coat them with the capture antibody (a detailed protocol is included). You will also need to prepare the various PBS buffers used during the ELISA assay. There will be enough reagents provided with the kit to run five 96-well plates.

With the ELISA kit ab46087 however, the buffers will be provided and a pre-coated 96-well plate will be included with the kit.

For the MDA kit, unfortunately we can not provide this lysis buffer as a separate component and it is only available as part of this kit.

I hope this information will be useful, but please let me know if you have further questions and I'll be happy to help.

Have a nice day.

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Answer

Thanks for your enquiry. Please see below for answers to your questions:

Response to question 1: No, 24 hr drying time is not necessary. After tapping the plate to remove excess liquid from the wells, it should be totally dry much faster than 24 hrs. Probably 30-60 min should be enough time. You can use a clean pipette tip to carefully remove excess liquid from the bottom of the wells if this will help things go faster.

Response to question 2: The antibody is stable for 6 monhts at 4 deg C. It does not need to be frozen.

Response to question 3: The kit has not been tested in the manner you describe. It is difficult to predictif the kit will peform as expected in this alternative assay format.Sorry I can't be more helpful on this one.

I hope this is helpful. Please contact me agaiin if you have any further questions.

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