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    mouse-tnf-alpha-elisa-kit-ab108910.pdf

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Immunology Innate Immunity Cytokines Necrosis Factors
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Mouse TNF alpha ELISA Kit (ab108910)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (3)References (3)

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Typical Standard Curve

    Key features and details

    • Sensitivity: 30 pg/ml
    • Range: 0.031 ng/ml - 2 ng/ml
    • Sample type: Cell culture supernatant, Plasma, Serum
    • Detection method: Colorimetric
    • Assay type: Sandwich (quantitative)
    • Reacts with: Mouse

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    Overview

    • Product name

      Mouse TNF alpha ELISA Kit
      See all TNF alpha kits
    • Detection method

      Colorimetric
    • Precision

      Intra-assay
      Sample n Mean SD CV%
      Overall 4.5%
      Inter-assay
      Sample n Mean SD CV%
      Overall 7.1%
    • Sample type

      Cell culture supernatant, Serum, Plasma
    • Assay type

      Sandwich (quantitative)
    • Sensitivity

      30 pg/ml
    • Range

      0.031 ng/ml - 2 ng/ml
    • Recovery

      97 %

    • Assay time

      5h 0m
    • Assay duration

      Multiple steps standard assay
    • Species reactivity

      Reacts with: Mouse
    • Product overview

      Abcam’s TNF alpha Mouse in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TNF alpha levels in plasma, serum and cell culture supernatants.


      A TNF alpha specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently a TNF alpha specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Conjugate is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize Streptavidin-Peroxidase enzymatic reaction. TMB is catalyzed by Streptavidin-Peroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the amount of TNF alpha captured in plate.


      The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.

    • Platform

      Microplate

    Properties

    • Storage instructions

      Store at -20°C. Please refer to protocols.
    • Components 1 x 96 tests
      100X Streptavidin-Peroxidase Conjugate 1 x 80µl
      10X Diluent N Concentrate 1 x 30ml
      1X Standard Diluent 1 x 2ml
      20X Wash Buffer Concentrate 2 x 30ml
      40X Biotinylated Mouse TNF alpha Antibody 1 x 150µl
      Chromogen Substrate 1 x 7ml
      Sealing Tapes 3 units
      Stop Solution 1 x 11ml
      TNF alpha Microplate (12 x 8 well strips) 1 unit
      TNF alpha Standard 1 vial
    • Research areas

      • Immunology
      • Innate Immunity
      • Cytokines
      • Necrosis Factors
      • Immunology
      • Innate Immunity
      • Cytokines
      • TNF Superfamily
      • Signal Transduction
      • Growth Factors/Hormones
      • TNF
      • Stem Cells
      • Signaling Pathways
      • TGF beta
      • TNF alpha
      • Cancer
      • Growth factors
      • TNF
      • Cancer
      • Tumor immunology
      • Cytokines
      • TNF
      • Cardiovascular
      • Atherosclerosis
      • Vascular Inflammation
      • Inflammatory mediators
      • Kits/ Lysates/ Other
      • Kits
      • ELISA Kits
      • ELISA Kits
      • TNF ELISA kits
      • Metabolism
      • Types of disease
      • Metabolic disorders
    • 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
      • 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

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

      Associated products

        Images

        • Typical Standard Curve
          Typical Standard Curve

          Representative Standard Curve using ab108910

        Protocols

        • Protocol Booklet

        Click here to view the general protocols

        Datasheets and documents

        • Datasheet
        • SDS
      • References (3)

        Publishing research using ab108910? Please let us know so that we can cite the reference in this datasheet.

        ab108910 has been referenced in 3 publications.

        • Gao J  et al. Metformin protects against PM2.5-induced lung injury and cardiac dysfunction independent of AMP-activated protein kinase a2. Redox Biol 28:101345 (2020). PubMed: 31669973
        • Wang H  et al. AMPKa2 deficiency exacerbates long-term PM2.5 exposure-induced lung injury and cardiac dysfunction. Free Radic Biol Med 121:202-214 (2018). PubMed: 29753072
        • Zheng J  et al. Maternal Low-Protein Diet Modulates Glucose Metabolism and Hepatic MicroRNAs Expression in the Early Life of Offspring †. Nutrients 9:N/A (2017). ELISA ; Mouse . PubMed: 28264458

        Customer reviews and Q&As

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

        1-3 of 3 Abreviews or Q&A

        Question

        We used RIPA lysis buffer and wondered whether it will have an effect on results. The RIPA buffer includes 150mM NaCl, 0.1% Triton X-100, 0.1% SDS, 0.5% sodium deoxycholate, and 50mM Tris-HCl. Another question that we have is that we used the Bradford Assay to determine our protein lysate concentrations. Do you think the protein concentrations determined from the Bradford is accurate enough, i.e. is there a better method that you would recommend to determine protein concentrations?

        Read More

        Abcam community

        Verified customer

        Asked on Nov 18 2013

        Answer

        The ELISA assays are compatible with RIPA buffer. However, you should prepare your standards and positive controls in the same buffer.

        The Bradford assay is compatible with RIPA buffer if the samples are diluted at least 10-fold with distilled water. Thermo Scientific/Pierce has a useful guide to protein assays in PDF form at the following link. http://www.piercenet.com/files/1602063_PAssayHB_122910.pdf . On page 12-13 you will see a table that compares the compatibility of various protein assays with many reagents, including RIPA buffer. The table refers to the Bradford assay as "Coomassie", an alternative name. The BCA assay is considered to be a better choice than the Bradford assay for assays of samples that contain detergents, but if you dilute your samples before assaying them, the Bradford may be adequate.

        Read More

        Tom Ruyle

        Abcam Scientific Support

        Answered on Nov 18 2013

        Question

        Hello, I am using ELISA kit number ab108910. I have an older plate reader on an older computer, and I cannot change the program settings for the reader. This ELISA kit comes with a white tray with clear strips of tubes, but I cannot change the setting on the plate reader to read a white plate with a clear bottom. I also cannot find any black tray that your ELISA strips fit in. Do you have black trays that your tube strips will fit into? Or can you recommend where I would find a black tray for the plate reader that your ELISA strips fit in?

        Read More

        Abcam community

        Verified customer

        Asked on Jan 28 2013

        Answer

        Thank you for contacting us.

        Our kit is color metric not chemoluminescent or fluorescent format. Only those format come with black frame. I don’t think the frame is an issue for the reader.


        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 Jan 28 2013

        Question

        Hello, Do you have a protocol for culturing cells to use the supernant for ELISAs? I am using your kits (ab108910 and ab108870) to detect TNFa and IL-10 in the supernatant from mouse macrophages, but I am having  (excel file attached). Any advice would be appreciated.

        Read More

        Abcam community

        Verified customer

        Asked on Nov 29 2012

        Answer

        Thank you for contacting us.

        I am very sorry to hear that you have been having troubles when using this product. In order that I might more fully understand the problem could you please tell me what is sample dilution factor was that you used? Sample concentration, both too high or too low can often, can lead to problems such as we see here. Next, how where your cell culture media samples collected and prepared? Were all reagents at room temperature before use?



        This product is covered by our Abpromise guarantee. We are happy provide scientific support at any time. If you are using the product in species and applications listed on the datasheet and contact us within six months of purchase, we are also happy to replace or refund the product should we not be able to help you to resolve the issue. More information on our Abpromise may be found at the following link:
        https://www.abcam.com/index.html?pageconfig=abpromise
        I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.





        Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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        Abcam Scientific Support

        Answered on Nov 29 2012

        Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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