製品の概要

  • 製品名

    Mouse TNF alpha ELISA Kit
    TNF alpha キット 製品一覧
  • 検出方法

    Colorimetric
  • 再現性

    Intra-Assay(同時再現性)
    サンプル N 平均値 SD CV%
    Overall 8 6.7%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    Overall 3 9.8%
  • サンプルの種類

    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • アッセイタイプ

    Sandwich (quantitative)
  • 検出感度

    9.1 pg/ml
  • 検出範囲

    46.88 pg/ml - 3000 pg/ml
  • 添加回収試験

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    Serum 97 85% - 115%
    Cell culture media 111 109% - 115%
    Heparin Plasma 92 85% - 103%
    EDTA Plasma 99 98% - 101%
    Citrate Plasma 87 85% - 88%

  • 全工程の試験時間

    1h 30m
  • ステップ

    One step assay
  • 種交差性

    交差種: Mouse
  • 製品の概要

    TNF alpha in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit (ab208348) is designed for the quantitative measurement of TNF alpha protein in mouse serum, plasma, and cell culture supernatants.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:


    Samples diluted in Sample Diluent NS - 9.8 pg/mL


    Samples diluted in Sample Diluent 10BS - 12.1 pg/mL


    Samples diluted in Sample Diluent 25BS - 9.1 pg/mL

  • 特記事項

    Tumor necrosis factor, or TNF alpha, is a cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. Furthermore, TNF alpha is a ligand of the TNF superfamily which plays a central role in inflammation, apoptosis, proliferation, invasion, angiogenesis, metastasis, and morphogenesis. It is mainly secreted by macrophages, and can induce cell death of certain tumor cell lines. Rat and Human TNF alpha are 95% and 79% identical to Mouse TNF alpha, respectively.

    TNF alpha is expressed on macrophages and endothelial, epithelial, and tumor cells as a 26 kDa transmembrane protein. TNF-alpha is cleaved by proteolytic processing into six chains: (1) TNF membrane form, (2) Intracellular domain 1 (ICD1), (3) Intracellular domain 2 (ICD2), (4) C-domain 1, (5) C-domain 2 and (6) TNF soluble form. ICD1 and ICD2 are released into the cytosol, while C-domain 1 and C-domain 2 are released into the extracellular space. Furthermore, 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.

    Signaling from TNF-alpha differs depending on the type of ligand initiating the signaling event (intracellular, membrane, or soluble). As an example, the membrane form of TNF-alpha appears to mediate anti-tumorigenic therapeutic responses whereas the soluble ligand is linked to inflammation and proliferation. Likewise, the TNF intracellular domain (ICD) form induces IL-12 production in dendritic cells.

  • アプリケーション

    適用あり: Sandwich ELISAmore details
  • 試験プラットフォーム

    Pre-coated microplate (12 x 8 well strips)

製品の特性

  • 保存方法

    Store at +4°C. Please refer to protocols.
  • 内容 1 x 96 tests
    10X Mouse TNF alpha Capture Antibody 1 x 600µl
    10X Mouse TNF alpha Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Mouse TNF alpha Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NBS 1 x 20ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 x 96 tests
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • 研究分野

  • 機能

    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.
  • 関連疾患

    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).
  • 配列類似性

    Belongs to the tumor necrosis factor family.
  • 翻訳後修飾

    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.
  • 細胞内局在

    Secreted and Cell membrane.
  • Information by UniProt
  • 別名

    • 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
  • 参照データベース

アプリケーション

Our Abpromise guarantee covers the use of ab208348 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Sandwich ELISA Use at an assay dependent concentration.

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  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curves and corrected for sample dilution. Undiluted samples are as follows: plasma (citrate) 100%, plasma (EDTA) 100%, and plasma (heparin) 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 1.68 ng/mL in plasma (citrate), 1.81 ng/mL in plasma (EDTA) and 1.47 ng/mL in plasma (heparin).

  • The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 100%, cell culture 100%, and RAW 264.7 LPS stimulated cell culture supernatants 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 1.63 ng/mL in serum, 1.00 ng/mL in cell culture and 6.07 ng/mL in RAW 264.7 LPS stimulated cell culture supernatants.

  • The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curves and corrected for sample dilution. Undiluted samples are as follows: RAW 264.7 LPS stimulated supernatant 50% and RAW 264.7 LPS unstimulated supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 6346.4 pg/mL in RAW 264.7 LPS stimulated supernatant and 83.7 pg/mL in RAW 264.7 LPS unstimulated supernatant. RAW 264.7 cells were cultured in HGDMEM with 100 μg/mL Kanamycin and 2 mM L-glutamine. Cells were starved for 24 hours and treated in the presence and absence of 5 μg/mL of LPS.

プロトコール

参考文献

This product has been referenced in:

  • Li Y  et al. Ischemia-induced ACSL4 activation contributes to ferroptosis-mediated tissue injury in intestinal ischemia/reperfusion. Cell Death Differ N/A:N/A (2019). Read more (PubMed: 30737476) »
  • Wu P  et al. MicroRNA-494-3p protects rat cardiomyocytes against septic shock via PTEN. Exp Ther Med 17:1706-1716 (2019). Read more (PubMed: 30783439) »
See all 26 Publications for this product

レビューと Q&A

1-2 of 2 Abreviews or Q&A

TNFa ELISA on mouse serum

Good Good 4/5 (Ease of Use)
Abreviews
We ran this kit on mouse serum pools from C57BL6 mice and did not get any meaningful signal and it was off the curve. Do not run on serum of mice that are not chronically inflamed or are not treated with something that would lead to a spike in TNFa. Other than that, a good ELISA, standard curve looked good.

Mr. Joshua Preston

Verified customer

投稿 Jun 26 2019

TNF alpha simple step ELISA kit

Good Excellent 5/5 (Ease of Use)
Abreviews
Good one and easy to use

Miss. Sheelu Monga

Verified customer

投稿 Feb 06 2018

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