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  1. Link

    products/elisa/rat-tnf-alpha-elisa-kit-ab100785.pdf

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Immunology Innate Immunity Cytokines Necrosis Factors
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Rat TNF alpha ELISA Kit (ab100785)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (5)References (77)

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Sandwich ELISA - TNF alpha Rat ELISA Kit (ab100785)
  • Sandwich ELISA - TNF alpha Rat ELISA Kit (ab100785)
  • Typical Standard Curve

Key features and details

  • Sensitivity: 25 pg/ml
  • Range: 82.3 pg/ml - 20000 pg/ml
  • Sample type: Cell culture extracts, Cell culture supernatant, Tissue Extracts
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Rat

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Overview

  • Product name

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

    Colorimetric
  • Sample type

    Cell culture supernatant, Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 25 pg/ml
  • Range

    82.3 pg/ml - 20000 pg/ml
  • Recovery

    92 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 93.17 81% - 105%
    Tissue Extracts 92.48 80% - 104%
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Rat
  • Product overview

    Abcam’s TNF alpha ELSA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Rat TNF alpha in cell lysates and tissue lysates.

    This assay employs an antibody specific for Rat TNF alpha coated on a 96-well plate. Standards and samples are pipetted into the wells and TNF alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Rat TNF alpha antibody is added. After washing away unbound biotinylated antibody, HRPconjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TNF alpha bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    200X HRP-Streptavidin Concentrate 1 x 200µl
    20X Wash Buffer 1 x 25ml
    2X Cell Lysis Buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    5X Sample Diluent Buffer 1 x 10ml
    Biotinylated anti-Rat TNF alpha 2 vials
    Recombinant Rat TNF alpha Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
    TNF alpha Microplate (12 x 8 wells) 1 unit
  • 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
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • 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
    • Neuroscience
    • Development
    • Neuroscience
    • Diseases
    • Neuroscience
    • Processes
  • 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: 24835 Rat
    • SwissProt: P16599 Rat
    • Unigene: 2275 Rat

    Images

    • Sandwich ELISA - TNF alpha Rat ELISA Kit (ab100785)
      Sandwich ELISA - TNF alpha Rat ELISA Kit (ab100785)

      Rat TNF alpha detected in cell lysates from NR8383.1 control cells (C) or cells stimulated for 6 hours with 5 ug x mL-1 of LPS (Sigma). Results shown for TNF alpha levels in 10e6 cells after background signal was subtracted (duplicates +/- SD).

    • Sandwich ELISA - TNF alpha Rat ELISA Kit (ab100785)
      Sandwich ELISA - TNF alpha Rat ELISA Kit (ab100785)

      Rat TNF alpha detected in cell supernatants from NR8383.1 control cells (C) or cells stimulated for 6 hours with 5 ug x mL-1 of LPS (Sigma); background signal subtracted (duplicates +/- SD).

    • Typical Standard Curve
      Typical Standard Curve

      Representative standard curve using ab100785

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (77)

    Publishing research using ab100785? Please let us know so that we can cite the reference in this datasheet.

    ab100785 has been referenced in 77 publications.

    • Walladbegi J  et al. Moderate temperature reduction is sufficient for prevention of 5-fluorouracil-induced oral mucositis: an experimental in vivo study in rats. Cancer Chemother Pharmacol 91:67-75 (2023). PubMed: 36451020
    • Zhao Y  et al. Hydrogen Sulfide Attenuates Lipopolysaccharide-Induced Inflammation via the P-glycoprotein and NF-κB Pathway in Astrocytes. Neurochem Res 48:1424-1437 (2023). PubMed: 36482035
    • Li L  et al. A nanofiber-hydrogel composite improves tissue repair in a rat model of Crohn's disease perianal fistulas. Sci Adv 9:eade1067 (2023). PubMed: 36598982
    • Phothi T  et al. Effects of curcumin and ?-oryzanol solid dispersion on the brain of middle-aged rats. Biomed Rep 17:59 (2022). PubMed: 35719843
    • Manzano-Pech L  et al. Excessive Consumption Hibiscus sabdariffa L. Increases Inflammation and Blood Pressure in Male Wistar Rats via High Antioxidant Capacity: The Preliminary Findings. Cells 11:N/A (2022). PubMed: 36139349
    View all Publications for this product

    Customer reviews and Q&As

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

    1-7 of 7 Abreviews or Q&A

    Testing TNF-alpha levels in rat spinal cord.

    Below average Good 4/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We tested TNF-α levels in homogenized rat spinal cord. The standard curve was good (R^2 = 0.999), but the sample results had to high of background and the CV values were extremely high. The tech help from ABCAM was very helpful. We tried several of their recommendations including using a blocker, different dilutions, centrifuging again before adding samples, and triplicate wells, but nothing helped.

    Note in the image high CV levels and the readings were just as high in wells where we did not add the detection antibody (ex. sample "B" is with detection antibody and sample "BND" is the same sample without detection antibody).
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 16 2020

    Abreview for TNF alpha Rat ELISA Kit

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image

    Sample: Rat Tissue lysate - whole (spinal cord)
    Specification: spinal cord
    Amount of Sample: 100 µg
    Type of Kit Used: ELISA
    Positive control: Traumatized segment level T13
    Did you use the Abcam Kit protocol?: Yes

    Additional data
    Additional Notes: I have used the kit with fresh rat spinal cord samples that underwent compression. The samples where homogenized a day before and kept in -20c. The strips allowed me to use the kit 3 times. The third time was a week after I have open Item F and still got a nice curve. R-square=0.9959.
    Results are more than satisfying. It is a quick and straight forward assay.

    Graph labels: Y axis is absorbance, X axis is pg/ml.The key labels are, from top to bottom: Standards, suppresses stds, standard curve.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Prof. Guy Sovak

    Verified customer

    Submitted May 21 2012

    Question

    I´m interested to measure IL-1 beta (Rat IL-1 beta ELISA Kit (Interleukin-1 beta) (ab100768)) and TNF (Rat TNF alpha ELISA Kit (ab100785) levels in rat hippocampus tissue. I've read the manual and it says that the kit can measure these cytokines levels (tissue homogenates),however it is not specified in what kind of buffer it must be homegenize the tissues. My question is: Can I use these Cytokines kits using homgenates in Lysis buffer ( 150 mM sodium chloride, 1.0% Triton X-100, 50 mM Tris pH 8.0)?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 16 2017

    Answer

    Yes, that lysis buffer will be good, the 150 mM sodium chloride, 1.0% Triton X-100, 50 mM Tris pH 8.0.


    Here are some general guidelines.


    Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as CHAPS, or mild ionic detergents such as sodium deoxycholate will work. 


    Use no more than 2% v/v total detergent  


    Avoid the use of sodium azide 


    Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols 


    We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to homogenization.

    Read More

    Abcam Scientific Support

    Answered on Jan 16 2017

    Question

    Does the cell lysate buffer contain any protease inhibitor?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 26 2014

    Answer

    Thank you for contacting us. The cell lysis buffer does not contain any protease inhibitor. Please let me know if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Nov 26 2014

    Question

    We would like to assess both IL-1 Beta and TNF alpha concentration in homogenate of colonic tissue, but in the data sheet is not specified the procedure to obtain the tissue homogenate. Do you have available a standardized extraction methodology? or could you suggest any references describing such a procedures?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 12 2014

    Answer



    I can suggest the following general tissue lysate preparation procedure to prepare samples for both the kits.




    Tissue lyste samples can be prepared using most conventional methods, for example with RIPA or other formulations.

    Please note the following guidelines on lysis buffer composition:
    1) Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic
    detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as
    CHAPS, or mild ionic detergents such as sodium deoxycholate will work.
    2) Use no more than 2% v/v total detergent
    3) Avoid the use of sodium azide
    4) Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols

    We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to
    homogenization. Most general biochemical supply companies including stock a wide variety of these products. Since susceptibility to proteolytic cleavage and the type of proteases present in the lysate vary, we do not recommend a specific product. Instead, your choice of which combination of protease inhibitors to use should be based upon a literature search for your protein(s) of interest and/or tissue or cell type. Phosphatase inhibitors may be used but are not necessary unless the antibodies used in the kit specifically
    recognize phosphorylated forms of the protein.
    Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freezethaw,
    sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these.
    There is no best method for all sample types; your choice of method should be made following a brief
    search of the literature to see how samples similar to yours have been prepared in previous
    investigations.
    After homogenization, centrifuge the lysates to remove cell/tissue debris (5 min @ 10,000 x g or 10
    min @ 5,000 x g) and save the supernatant. Unless testing fresh, lysates should be frozen as soon as
    possible and stored at -20°C (or -80°C, if possible). Centrifuge them again before incubating with any
    immunoassay. Next, determine the protein concentration of your lysates using a total protein assay
    not inhibited by detergents (such as the Bicinchoninic acid (BCA) assay) and normalize the volume of
    each sample used to deliver the same amount of total protein for each assay.

    Note: The Bradford assay is not recommended as it can be inhibited by the presence of detergents.
    Since different cells and tissues may contain different amounts of protein, as starting point, we
    suggest using 500 μL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust this
    based upon your results. Your target total protein concentration of the homogenate should be at least
    1,000 μg/mL, but 2,000 μg/mL or more would be better.

    Read More

    Sam Washer

    Abcam Scientific Support

    Answered on Mar 12 2014

    Question

    Does this ab100785 product contain any hazardous components for Japan customers?

    Read More

    Abcam community

    Verified customer

    Asked on May 25 2012

    Answer

    Thank you for your enquiry.

    Please rest assured that there are no hazardous components in this kit according to Japanese law, and is safe for use by customers in Japan.

    Have a nice day!

    Read More

    Abcam Scientific Support

    Answered on May 25 2012

    Question

    What is Item J (Cell lysate buffer) for? How much sample should I prepare?

    Read More

    Abcam community

    Verified customer

    Asked on May 14 2012

    Answer

    Thank you for your enquiry.

    The lysis buffer (item J) is used to prepare the cell and tissue sample. There is no mention on how to prepare your sample in the Protocol Booklet as most laboratories would have their own protocols and methods for doing so. We do recommend starting with 1mg/ml, preferable 2mg/ml volume (sample in lysis buffer) of sample. Please bear in mind that this is a starting point and the optimal concentration needs to be determined empirically.

    You may also use your own lysis buffer. As a guideline, a lysis buffer must meet the following specifications.

    A) has relatively low salt content (700 mM or less)
    B) does not contain sodium azide
    C) does not contain >0.1% SDS
    D) does not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol)

    I hope this information will be helpful.

    Read More

    Abcam Scientific Support

    Answered on May 14 2012

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    For licensing inquiries, please contact partnerships@abcam.com

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