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

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • 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, macrophage derived
    • TNF, monocyte derived
    • TNF-a
    • TNF-alpha
    • TNFA
    • TNFA_HUMAN
    • TNFSF2
    • Tumor necrosis factor
    • 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

Applications

Our Abpromise guarantee covers the use of ab46070 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

  • 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).

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

  • Representative Standard Curve using ab46070

Protocols

References

This product has been referenced in:
  • Jiang P  et al. Astragaloside IV Prevents Obesity-Associated Hypertension by Improving Pro-Inflammatory Reaction and Leptin Resistance. Mol Cells 41:244-255 (2018). Read more (PubMed: 29562733) »
  • Gu R  et al. A Synthesized Glucocorticoid- Induced Leucine Zipper Peptide Inhibits Retinal Müller Cell Gliosis. Front Pharmacol 9:331 (2018). Read more (PubMed: 29681857) »
See all 21 Publications for this product

Customer reviews and Q&As

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

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

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.



Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

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Question
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

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

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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