概述

  • 产品名称
    人TNF alpha ELISA试剂盒
    参阅全部 TNF alpha 试剂盒
  • 检测方法
    Colorimetric
  • 精确度
    批次内
    样品 n Mean SD CV%
    PBMC media 8 2.3%
    批次间
    样品 n Mean SD CV%
    PBMC media 3 5%
  • 样品类型
    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • 检测类型
    Sandwich (quantitative)
  • 灵敏度
    14 pg/ml
  • 范围
    31.25 pg/ml - 2000 pg/ml
  • 回收率

    特定样本回收率
    样品类型 平均% 范围
    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%

  • 检测时间
    1h 30m
  • 实验步骤
    One step assay
  • 种属反应性
    与反应: Human
    不与反应: Mouse
  • 产品概述

    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.

  • 说明

    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.

  • 经测试应用
    适用于: Sandwich ELISAmore details
  • 平台
    Microplate

性能

  • 存放说明
    Store at +4°C. Please refer to protocols.
  • 组件 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    ab221825 - 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 Development Solution 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
  • 研究领域
  • 功能
    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.
  • 疾病相关
    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).
  • 序列相似性
    Belongs to the tumor necrosis factor family.
  • 翻译后修饰
    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.
  • 细胞定位
    Secreted and Cell membrane.
  • Information by UniProt
  • 别名
    • 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
  • 数据库链接

应用

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.

应用 Ab评论 说明
Sandwich ELISA Use at an assay dependent concentration.

图片

  • 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. 

实验方案

文献

This product has been referenced in:
  • Huang G  et al. Platelet-rich plasma shows beneficial effects for patients with knee osteoarthritis by suppressing inflammatory factors. Exp Ther Med 15:3096-3102 (2018). Read more (PubMed: 29599843) »
  • Moussa BA  et al. Synthesis and molecular docking of new roflumilast analogues as preferential-selective potent PDE-4B inhibitors with improved pharmacokinetic profile. Eur J Med Chem 148:477-486 (2018). Read more (PubMed: 29477888) »

See all 12 Publications for this product

客户评价及客户问答

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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提交于 Jun 01 2016

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