Rat TNF alpha ELISA Kit (ab46070)

Reviews (1)Q&A (5)References (46)

Overview

  • Product name

    Rat TNF alpha ELISA Kit
    See all TNF alpha kits

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    A 12 543.5pg/ml 44.7 8.2%
    B 10 130.1pg/ml 10.3 7.9%
    Inter-assay
    Sample n Mean SD CV%
    A 9 541.4pg/ml 33.2 6.1%
    B 9 149.4pg/ml 9.8 6.5%

  • Sample type

    Cell culture supernatant, Serum

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    < 15 pg/ml

  • Range

    31.25 pg/ml - 1000 pg/ml

  • Assay time

    3h 45m

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Rat

  • Product overview

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


    A monoclonal antibody specific for TNF alpha has been coated onto the wells of the microtiter strips provided. Samples, including standards of known TNF alpha concentrations, control specimens or unknowns are pipetted into these wells. During the first incubation, the standards or samples and a biotinylated monoclonal antibody specific for TNF alpha are simultaneously incubated. After washing, the enzyme Streptavidin-HRP, that binds the biotinylated antibody is added, incubated and washed. A TMB substrate solution is added which acts on the bound enzyme to induce a colored reaction product. The intensity of this colored product is directly proportional to the concentration of TNF alpha present in the samples.


    This kit will recognize both endogenous and recombinant rat TNF alpha

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components Identifier 1 x 96 tests 2 x 96 tests
    10X Standard Diluent Buffer Black 1 x 25ml 1 x 25ml
    200X Wash Buffer White 1 x 10ml 2 x 10ml
    Biotinylated Antibody Diluent Red 1 x 7.5ml 1 x 13ml
    Biotinylated anti-TNF alpha Red 1 x 400µl 2 x 400µl
    Chromogen TMB Substrate Solution 1 x 11ml 1 x 24ml
    HRP Diluent Red 1 x 23ml 1 x 23ml
    Plastic Plate Covers 2 units 4 units
    Stop Reagent Black 1 x 11ml 2 x 11ml
    Streptavidin-HRP 2 x 5µl 4 x 5µl
    TNF alpha Microplate (12 x 8 well strips) 1 unit 2 units
    TNF alpha Standard (Lyophilized) Yellow 2 vials 4 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

    Associated products

    Images

    • ELISA: TNF alpha Rat ELISA Kit (ab46070)
      ELISA: TNF alpha Rat ELISA Kit (ab46070)

      Rat TNF alpha measured in culture supernatants (dilution range 1/1-1/200) from control NR8383 cells or cells stimulated for 24 hours with 50 ng x mL-1 of PMA ((PMA) ab120297) or PMA with the addition of 1 ug x mL-1 of LPS (Sigma) (LPS+PMA) for the last 6 hours (duplicates +/- SD).

    • ELISA: TNF alpha Rat ELISA Kit (ab46070)
      ELISA: TNF alpha Rat ELISA Kit (ab46070)

      Standard curve with background signal subtracted (duplicates; +/- SD).

    • Typical Standard Curve
      Typical Standard Curve

      Representative Standard Curve using ab46070

    Protocols

    References

    This product has been referenced in:

    • Wang C  et al. Low-frequency pulsed electromagnetic field promotes functional recovery, reduces inflammation and oxidative stress, and enhances HSP70 expression following spinal cord injury. Mol Med Rep 19:1687-1693 (2019). Read more (PubMed: 30628673) »
    • Sheibani M  et al. Sumatriptan protects against myocardial ischaemia-reperfusion injury by inhibition of inflammation in rat model. Inflammopharmacology N/A:N/A (2019). Read more (PubMed: 30929154) »
    See all 46 Publications for this product

    Publishing research using ab46070? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-6 of 6 Abreviews or Q&A

    Question


    First of all, I want to thank you for your immediate and explicit explanations.
    I have requested the purchase date and order number of ab100786 VEGF
    and the others from the university. I will email them with a better
    resolution of the results to you ASAP I get access to them.
    For the TNF kit ab46070, as I have understood, it seems like nothing
    more can be done with the results. Please correct me if I am wrong.
    For the third kit, I am so so sorry, since a little misspelling I
    have done. The kit number is ab100772 IL6 rat, instead of 100722 !
    However, the file you have sent to me is not what I was expecting. I
    do have the brochure of the kit. My problem was something else. The
    standards did not work and so a curve was not drawn. I wanted you to
    email me, if possible offcourse, the optimized numbers for STANDARDs
    of this kit in your lab. I mean, I am aware that each kit&apos;s standards should be optimized in case, but since my standards did not give me concentrations, although I have OD of my samples, I cannot know the concentrations of IL6 in the samples. So if possible, could you provide me the optimized numbers for OD and concentrations of
    standards of this kit? (I mean the raw numbers, regardless of the fact
    that they should be assessed in each case separately). So I would be
    able to draw a curve by them and calculate the concentration of IL6 in
    my samples upon the OD that the plate reader has provided.
    Here, I would like to express my gratitudes for you co-operation.
    Kind Regards,
    Afshin Azimirad.

    Read More
    Answer

    Thank you for your reply.

    I look forward to receiving the resuls and order number for the ab100786 VEGF rat ELISA kit.

    For the TNF kit ab46070, I think it is likely to be the sample that is the problem.However, I would be happy to investigate this further. In order to do so, I would appreciate if you are able to please complete the questionaire below and also provide the order number, lot number and a copy of the results (including the standard curve). As discussed in the previous email, it is difficult to make suggestions without haveing furhter details, and so this information will be very helpful for our investigations for you. The kit is covered by our 6 month guarantee.

    With regards to ab100772IL6 Rat ELISA Kit, the standard curve using the mouse standard provided with the kit shouldbe working and we can provide our guarantee for this.I am forwarding the protocol file for this kit. I am sending this to confirm that the protocol should be undertaken exactly as described in the protocol provided with the kit,including thepreparation and detectionof the standard curve as well as the samples. Could you confirm this protocol has been followed?

    https://www.abcam.com/ps/products/100/ab100772/documents/ab100772-IL6-Rat-ELISA-Kit-ab100772-protocol-plain-v2.pdf

    I would appreciate if you are able to provide further information and theresults of the standard from this kit (ab100722) and I will be pleased to investigate further. In order to do so, I have attached a copy of a questionaire below. This will help you to put together the information we require quickly and easily.

    Thank you for your continued time and cooperation. I look forward to hearing from you.



    A. General Information
    - Catalogue number:


    - Lot number:

    - Purchase order number/ order date or preferably the Abcam order number:

    - How was the kit stored?


    B. Description of the problem
    - Are the signals you obtained with control or standard satisfactory? (Yes/No);


    - Are the sample readings as expected or inconsistent?


    - Are the blank readings as expected?

    - Other (e.g. missing components)


    C. What do you think the problem is caused by? E.g. enzyme is faulty, developer is faulty or other component that you think might be faulty.



    D. Sample
    - Sample Type (e.g. Cell culture supernatant, Cell lysates, Tissue lysates or Serum samples)


    - Sample Species (e.g. human, mouse or rat)


    - Storage (at what temperature and for how long were the samples stored?)

    - Sample treatment, if any



    - Sometime proteins in samples interfere with readings. Have you removed proteins?


    - Sample preparation


    E. Protocol used:
    Did you follow the protocol exactly as given on the datasheet?


    F. Optimization attempts (problem solving); please describe any deviations from the protocol.



    G. Have you used the same kit successfully before?


    H. Do you obtain the same results every time?


    I. Additional Notes; (any additional information you would like to provide)



    J. Results:

    Read More

    Question


    I am sorry bothering you again. I will sent you the numbers of the
    kit with a better image of the results soon. Meanwhile, I have
    consumed two other kits of your company and unfortunately I faced some
    problem with them,too.
    The second kit ab46070 TNF-alpha rat : I used this kit for
    peritoneal wash of my rats. The all numbers (OD & Quant) reported as
    zero. The standards were as they were supposed. Does it mean that this
    kit does not support peritoneal wash ? and just serum/plasma should be
    used?
    The third kit was an ab100722 kit for IL6 rat. The procedure was
    ok. However, the ELISA reader only reported ODs, and not the
    quantities. In fact, the standars are not curved. I am aware that
    every kit has its own standard ODs and numbers and it should be
    individualized. However, since the quantity results are important for
    me, may I favour you to email me the standard table for this kit,
    which has been optimized in your lab, regardless of the
    individualization. Hence, I might be able to interprete the OD s by
    some degrees.
    Here again, I should express my gratitude for your kind co-operation.
    Best Regards,

    Read More
    Answer

    Thank you for your message. I look forward to receiving the further information regarding ab100786 VEGF Rat ELISA Kitso we can investigate this further for you.

    I am sorry to hear you also have some concerns regarding two other kits. I would like to reassure you that these will also be covered by the 6 month guarantee.

    ab46070 TNF-alpha rat: if the standards were ok,this indicates that the sample does not contain enough TNF alpha for detection (perhaps it has become too dilute with the washing). Do you have a good positive control sample you could try? It is possible, although notas likely, that the kit is not compatible with this sample type, I am sorry this has not been tested. For our records, I would appreciate if you are able to provide the order number and date of purchase.

    ab100722 kit for IL6: I am sorry this has been tested in mouse, and not rat. I am sorry we would therefore not be able to guarantee the results from rat samples. However, the standard curve using the mouse standard provided with the kit should still be working and we can provide our guarantee for this. The protocol should be undertaken exactly as described in the protocol provided with the kit. Coud you confirm this?

    https://www.abcam.com/ps/products/100/ab100722/documents/ab100722-Mouse-MCP1-ELISA-Kit-ab100722-protocol-plain-v2.pdf

    I would appreciate if you are able to provide further information and the results of the standard from this kit (ab100722) and I will be pleased to investigate further. In order todo so, I have attached a copy of a questionaire below. This will help you to put together the information we require quickly andeasily.

    Thank you for your time and continued cooperation. I look forward to hearing from you with the requested information.




    A. General Information
    - Catalogue number:

    - Lot number:

    - Purchase order number/ order date or preferably the Abcam order number:

    - How was the kit stored?


    B. Description of the problem
    - Are the signals you obtained with control or standard satisfactory? (Yes/No);


    - Are the sample readings as expected or inconsistent?


    - Are the blank readings as expected?

    - Other (e.g. missing components)


    C. What do you think the problem is caused by? E.g. enzyme is faulty, developer is faulty or other component that you think might be faulty.



    D. Sample
    - Sample Type (e.g. Cell culture supernatant, Cell lysates, Tissue lysates or Serum samples)


    - Sample Species (e.g. human, mouse or rat)


    - Storage (at what temperature and for how long were the samples stored?)

    - Sample treatment, if any



    - Sometime proteins in samples interfere with readings. Have you removed proteins?


    - Sample preparation


    E. Protocol used:
    Did you follow the protocol exactly as given on the datasheet?


    F. Optimization attempts (problem solving); please describe any deviations from the protocol.



    G. Have you used the same kit successfully before?


    H. Do you obtain the same results every time?


    I. Additional Notes; (any additional information you would like to provide)



    J. Results:

    Read More

    Question

    Customer kindly called to inquire about the separate components used in this product. Specifically they wsh to be able to purchase another 96 well plate and to coat this with the antibody.

    Read More
    Answer

    Thank you for contacting us.

    I have been in contact with the production group regarding your inquiry. Unfortunately, at this time, we do not offer individual components of this kit for sale. Information about capture antibody used to coat the wells is also considered proprietary at this time and I am unable to obtain further information about this.

    I am sorry that I have been unable to provide the exact information you need but please do not hesitate to contact us if you need any more advice or information.



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    Question

    Gibt es das Streptavidin HRP separat?

    Read More
    Answer

    Vielen Dank für Ihre Anfrage.

    Wie versprochen habe ich mich mit dem Hersteller und unserer Produktabteilung in Verbindung gesetzt, um zu erfahren, ob es das Streptavidin-HRP separat erhältlich gibt. Leider haben wir diese Einzelkomponente von ab46070 derzeit nicht im Katalog.

    Ich möchte Ihnen jedoch gerne mitteilen, dass das Streptavidin-HRP für den Kit in einer Konzentration von 0.15 µg/ml eingesetzt wird. Streptavidin-HRP anderer Herkunft kann verwendet werden; eine Optimierung empfiehlt sich jedoch. Falls Sie in Ihrem Labor grade kein Streptavidin-HRP zur Hand haben sollten, eignet sich vielleicht unser ab7403 (Click here (or use the following: https://www.abcam.com/index.html?datasheet=7403).)

    Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Question

    What is the minimum detectable concentration of TNF alpha? We want to be able to detect as little as 5 pg/ml.

    Read More
    Answer

    Thank you for contacting us. The limit of detection of the TNF alpha ELISA kit ab46070 is 15 pg/ml, which is arrived at by taking the mean optical density of 64 blank wells (no TNF alpha), adding two standard deviations of the mean, then calculating the corresponding concentration from the standard curve. Simply including one or more standard dilutions will not improve the sensitivity. 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

    Abreview for TNF alpha Rat ELISA Kit

     Good Good 4/5 (Ease of Use)
    Abreviews
    Sample: Rat Serum (rat serum)
    Specification: rat serum
    Amount of Sample: 0.1 µg
    Type of Kit Used: ELISA
    Positive control: Serial dilution of target
    Did you use the Abcam Kit protocol?: Yes

    Additional data
    Additional Notes: Concentration estimated as serum was used.

    Mrs. Rhoda Cortes

    Verified customer

    Submitted Aug 30 2007

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