製品の概要

  • 製品名
    Human 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%
    PBMC media 8 2.3%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    PBMC media 3 5%
  • サンプルの種類
    Serum, Plasma, Cell culture media
  • アッセイタイプ
    Sandwich (quantitative)
  • 検出感度
    14 pg/ml
  • 検出範囲
    31.25 pg/ml - 2000 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 99.5 80% - 119%
    Cell culture media 100.97 95% - 109%
    Heparin Plasma 98 91% - 106%
    EDTA Plasma 102 95.7% - 110%
    Citrate Plasma 93.09 83.5% - 99%

  • 全工程の試験時間
    1h 30m
  • ステップ
    One step assay
  • 種交差性
    交差種: Human
    非交差種: Mouse
  • 製品の概要

    Abcam’s TNF-alpha in vitro SimpleStep ELISA®  (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TNF-alpha protein in Human serum, plasma and culture media.

    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.

  • 特記事項

    TNF-alpha, also known as cachectin or TNFSF1A, is the prototypic ligand of the TNF superfamily which plays a central role in inflammation, apoptosis, proliferation, invasion, angiogenesis, metastasis and morphogenesis. It is expressed on macrophages, endothelial, epithelial and tumor cells as a 26kDa transmembrane protein. TNF-alpha is cleaved by proteolytic processing into six chains: (1) TNF membrane form, (2) Intracellular domain 1, (3) Intracellular domain 2, (4) C-domain 1, (5) C-domain 2 and (6) TNF soluble form. 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.

  • 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
    Microplate

製品の特性

  • 保存方法
    Store at +4°C. Please refer to protocols.
  • 内容 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent 50BP 1 x 20ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
    TNFa Capture Antibody (Lyophilized) 1 x 2 vials
    TNFa Detector Antibody (Lyophilized) 1 x 2 vials
    TNFa Human Lyophilized Recombinant Protein 1 x 2 vials
  • 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 protein
    • Cachectin
    • DIF
    • soluble form
    • Tnf
    • TNF a
    • TNF macrophage derived
    • TNF monocyte derived
    • TNF superfamily member 2
    • TNF-a
    • TNF-alpha
    • TNFA_HUMAN
    • TNFSF2
    • Tumor necrosis factor
    • Tumor necrosis factor alpha
    • Tumor necrosis factor ligand superfamily member 2
    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 ab181421 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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  • Background-subtracted data values (mean +/- SD) are graphed. 

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

  • Human PBMCs were cultured in RPMI supplemented with 10% fetal calf serum, 2mM L-glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin.  Cells were cultured for 2 days at 37°C in the presence or absence of PHA.  The concentrations of TNF-alpha were interpolated from the calibration curve and corrected for sample dilution.  The mean TNF-alpha concentration was determined to be undetectable in basal media, 1.2 ng/mL in unstimulated PMBC supernatants and 4.7 ng/mL in stimulated PBMC supernatants. 

プロトコール

参考文献

This product has been referenced in:
  • Murphy RA  et al. Establishment of HK-2 Cells as a Relevant Model to Study Tenofovir-Induced Cytotoxicity. Int J Mol Sci 18:N/A (2017). ELISA ; Human . Read more (PubMed: 28257038) »
  • Tan SS  et al. Melatonin promotes survival of non-vascularized fat grafts and enhances the viability and migration of human adipose-derived stem cells via down-regulation of acute inflammatory cytokines. J Tissue Eng Regen Med N/A:N/A (2017). ELISA ; Human . Read more (PubMed: 28486770) »

See all 5 Publications for this product

レビューと Q&A

efficient and accurate

Excellent Excellent 5/5 (Ease of Use)
Abreviews
I used this kit to detect TNF-a in stimulated human CD14+ monocytes. The assay was extremely efficient. There is no overnight coating step, and detection/capture antibodies are added as a cocktail at the same time as the samples. The assay was quite accurate, detecting even minimal amounts of TNF-a. I would use this assay again for its efficiency, requiring very little time at the bench.
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投稿 Jun 01 2016

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

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