製品の概要

  • 製品名

    Human Thrombomodulin ELISA Kit
    Thrombomodulin 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%
    Human serum 8 3.1%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    Human serum 3 9.4%
  • サンプルの種類

    Cell culture supernatant, Urine, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma
  • アッセイタイプ

    Sandwich (quantitative)
  • 検出感度

    3 pg/ml
  • 検出範囲

    20.3 pg/ml - 1300 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
    Cell culture supernatant 90 88% - 91%
    Urine 106 102% - 112%
    Serum 97 96% - 99%
    Cell culture extracts 94 92% - 96%
    Tissue Extracts 105 101% - 110%
    Hep Plasma 99 98% - 100%
    EDTA Plasma 96 94% - 98%
    Cit plasma 96 92% - 98%

  • 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: Cow
  • 製品の概要

    Thrombomodulin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Thrombomodulin protein in serum, plasmas, urine, cell culture supernatants, and cell and tissue extract samples.


    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 in Sample Diluent NS: 11 pg/mL
    Samples in 1X Cell Extraction Buffer PTR: 3 pg/mL

  • 特記事項

    Human Thrombomodulin is a 557 amino acid type I transmembrane protein, consisting of six epidermal growth factor (EGF)-like domains and is highly glycosylated. Thrombomodulin is an important component in the anti-coagulation system by functioning as a cell surface receptor for thrombin, and has also been shown to play a role in anti-inflammation, cell adhesion, and proliferation. The Thrombomodulin - thrombin complex activates protein C and thrombin-activatable fibrinolysis inhibitor (TAFI); protein C degrades coagulation factors Va and VIIIa, which regulates thrombin resulting in inhibition of coagulation. The extracellular portion of Thrombomodulin has been shown to be released from the cell surface due to protein cleavage during vascular endothelial cell injury.

  • 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 Wash Buffer PT (ab206977) 1 x 20ml
    20X Human Thrombomodulin Detector Antibody 1 x 300µl
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    Human Thrombomodulin Capture Antibody (Lyophilized) 1 vial
    Human Thrombomodulin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    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
  • 研究分野

  • 機能

    Thrombomodulin is a specific endothelial cell receptor that forms a 1:1 stoichiometric complex with thrombin. This complex is responsible for the conversion of protein C to the activated protein C (protein Ca). Once evolved, protein Ca scissions the activated cofactors of the coagulation mechanism, factor Va and factor VIIIa, and thereby reduces the amount of thrombin generated.
  • 組織特異性

    Endothelial cells are unique in synthesizing thrombomodulin.
  • 関連疾患

    Defects in THBD are the cause of thrombophilia due to thrombomodulin defect (THR-THBD) [MIM:188040]. A hemostatic disorder characterized by a tendency to thrombosis.
    Defects in THBD are a cause of susceptibility to hemolytic uremic syndrome atypical type 6 (AHUS6) [MIM:612926]. An atypical form of hemolytic uremic syndrome. It is a complex genetic disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal failure and absence of episodes of enterocolitis and diarrhea. In contrast to typical hemolytic uremic syndrome, atypical forms have a poorer prognosis, with higher death rates and frequent progression to end-stage renal disease. Note=Susceptibility to the development of atypical hemolytic uremic syndrome can be conferred by mutations in various components of or regulatory factors in the complement cascade system. Other genes may play a role in modifying the phenotype.
  • 配列類似性

    Contains 1 C-type lectin domain.
    Contains 6 EGF-like domains.
  • 翻訳後修飾

    N-glycosylated.
    The iron and 2-oxoglutarate dependent 3-hydroxylation of aspartate and asparagine is (R) stereospecific within EGF domains.
  • 細胞内局在

    Membrane.
  • Information by UniProt
  • 別名

    • AHUS 6
    • AHUS6
    • BDCA 3
    • BDCA3
    • CD 141
    • CD141
    • CD141 antigen
    • Fetomodulin
    • Thbd
    • THPH12
    • THRM
    • Thrombomodulin
    • TM
    • TRBM_HUMAN
    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 ab214029 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.

画像

  • 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.

     

  • Standard curve comparison between human Thrombomodulin SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 8-fold increase in sensitivity.

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

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

  • The concentrations of Thrombomodulin were measured in duplicate, interpolated from the Thrombomodulin standard curve and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (citrate) 25%, plasma (EDTA) 25% and plasma (heparin) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombomodulin concentration was determined to be 4.8 ng/mL in serum, 3.9 ng/mL in plasma (citrate), 3.8 ng/mL in plasma (EDTA) and 4.3 ng/mL in plasma (heparin).

  • The concentrations of Thrombomodulin were measured in duplicate, interpolated from the Thrombomodulin standard curve and corrected for sample dilution. Undiluted sample is at 45%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The native Thrombomodulin concentration was undetectable in the null spike control run in parallel with the spiked sample.

  • The concentrations of Thrombomodulin were measured in duplicate and interpolated from the Thrombomodulin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombomodulin concentration was determined to be 1.2 ng/mL in HUVEC extract and 8.0 ng/mL in liver homogenate extract.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombomodulin concentration was determined to be 5.1 ng/mL with a range of 3.6 – 6.8 ng/mL.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean target concentration was determined to be 5.1 ng/mL with a range of 2.7 – 9.9 ng/mL.

プロトコール

参考文献

This product has been referenced in:

  • Kamtchum-Tatuene J  et al. HIV is associated with endothelial activation despite ART, in a sub-Saharan African setting. Neurol Neuroimmunol Neuroinflamm 6:e531 (2019). ELISA . Read more (PubMed: 30697583) »
See 1 Publication for this product

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