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Human Thrombin ELISA Kit (Factor II) (ab108909)

Reviews (1)Q&A (4)References (8)
Typical Standard Curve

    Key features and details

    • Sensitivity: 71 pg/ml
    • Range: 1.25 ng/ml - 10 ng/ml
    • Sample type: Cell culture supernatant, Cell Lysate, Tissue
    • Detection method: Colorimetric
    • Assay type: Sandwich (quantitative)
    • Reacts with: Human

    製品の概要

    • 製品名

      Human Thrombin ELISA Kit (Factor II)

    • 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 5.7%
      Inter-Assay（日差再現性）
      サンプル N 平均値 SD CV%
      Overall 10.1%

    • サンプルの種類

      Cell culture supernatant, Tissue, Cell Lysate

    • アッセイタイプ

      Sandwich (quantitative)

    • 検出感度

      = 71 pg/ml

    • 検出範囲

      1.25 ng/ml - 10 ng/ml

    • 添加回収試験

      98 %

    • 全工程の試験時間

      4h 0m

    • ステップ

      Multiple steps standard assay

    • 種交差性

      交差種: Human

    • 製品の概要

      Abcam’s Thrombin (Factor II) Human in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Thrombin concentrations in cell culture supernatants. This assay recognizes both natural and recombinant Human thrombin.


      A Thrombin specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently a Thrombin specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Conjugate is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize Streptavidin-Peroxidase enzymatic reaction. TMB is catalyzed by Streptavidin-Peroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the amount of Thrombin captured in plate.


      The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.

    • 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

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab108909の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

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

    画像

    • Typical Standard Curve
      Typical Standard Curve

      Representative Standard Curve Using ab108909.

    参考文献 (8)

    ab108909 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab108909 は 8 報の論文で使用されています。

    • Takayama W  et al. Dielectric Blood Coagulometry for the Early Detection of Sepsis-Induced Disseminated Intravascular Coagulation: A Prospective Observational Study. Crit Care Med 50:e31-e39 (2022). PubMed: 34369427
    • Feser CJ  et al. Engineering CRISPR/Cas9 for Multiplexed Recombinant Coagulation Factor Production. Int J Mol Sci 23:N/A (2022). PubMed: 35563479
    • Lei J  et al. Dabigatran activates inflammation resolution by promoting fibrinogen-like protein 2 shedding and RvD5n-3 DPA production. Theranostics 11:4251-4261 (2021). PubMed: 33754059
    • Zitur LJ  et al. Brain Death Enhances Activation of the Innate Immune System and Leads to Reduced Renal Metabolic Gene Expression. Transplantation 103:1821-1833 (2019). PubMed: 30964836
    • Wang H  et al. N-glycosylation in the protease domain of trypsin-like serine proteases mediates calnexin-assisted protein folding. Elife 7:N/A (2018). PubMed: 29889025
    • Moñux G  et al. FXa inhibition by rivaroxaban modifies mechanisms associated with the pathogenesis of human abdominal aortic aneurysms. Br J Clin Pharmacol 83:2661-2670 (2017). PubMed: 28735510
    • Abu El-Asrar AM  et al. Upregulation of Thrombin/Matrix Metalloproteinase-1/Protease-Activated Receptor-1 Chain in Proliferative Diabetic Retinopathy. Curr Eye Res 41:1590-1600 (2016). PubMed: 27261371
    • Liu JF  et al. Thrombin induces heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways. Arthritis Res Ther 14:R91 (2012). Sandwich ELISA ; Human . PubMed: 22541814

    レビューと Q&A

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    1-5 of 5 Abreviews or Q&A

    Testing Thrombin Human ELISA Kit for crossreactivity with rhesus plasma and sera:

     Good Excellent 5/5 (Ease of Use)
    Abreviews
    abreview image
    The thrombin ELISA, using the Abcam human ELISA kit Ab108909, lot GR232366-8, was performed by an experienced laboratory technician at an accredited university. The goal of the experiment was to analyze the linearity of dilutions of rhesus macaque EDTA-prepared plasma samples and serum samples.

    Overall, the kit, like other Abcam ELISA kits, was easy to follow, clear in terms of directions, and brief yet thorough. The laboratory technician had no problems understanding any of the protocol’s steps.

    Our test was performed in 2 basic steps. The first was to find the appropriate testable dilution in plasma and serum, and the second was to test the linearity and spike recovery.

    To this end our first experiment tested 8 dilutions of both serum and plasma ranging from 1:10 to 1:163840 (8 1:4 serial dilutions). The plasma was not in range of the assay, confirming that his kit does not detect pro-thrombin. The serum dilutions gave OD values in range of the assay from 1:10 – 1:2560, but a linear range was not observed.

    We saved the thrombin standard vial at -80C for 1 week between tests.

    Our second experiment consisted of a narrower dilution range at 1:50, 1:100, 1:200, 1:400,a nd 1:800 for n=2 rhesus sera and a spike recovery test. The tested range was well within the OD limits of the assay. N=1 rhesus serum may have been near linear range at 1:400-1:800, but n=1 rhesus serum was not. The spike recovery (performed with Serum from animal 2) showed an increasing spike recovery with dilution (40% at 1:50, 80% at 1:800).

    Overall, we would recommend this kit for its ease of use. We believe there is positive cross-reactivity for rhesus, but we were unable to identify a linear range for the assay.
    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

    投稿 Jun 07 2016

    Question

    I received the abcam Thrombin Elisa Assay today. In anticipation of my experiment next week, I wanted to make the wash buffer solution. Instead of adding the 20X wash buffer concentrate to the dH2O, I added 10 mLs of the 10X diluent instead. I wanted to know what the diluent buffer components are so if I need more, I can make it.

    Read More
    Answer

    Thank you for your patience while I looked into this.

    The lab has sent us thereplacement vials of the diluent and wash buffer. We are sending these to you free of charge on order. You should receive these tomorrow.

    I hope this information helps. Please contact us with any other questions.

    Read More

    Question

    How shall I reconstitute the standard? The lot number is GR88036-1.

    Read More
    Answer

    Thank you for contacting Abcam regarding ab108909.


    Just to confirm what we discussed over the phone:


    Thrombin Standard: Purified human Thrombin in a buffered protein base (40 ng, lyophilized).

    Standard Curve: Reconstitute the 40 ng of human Thrombin Standard with 2 ml of EIA Diluent to generate 20 ng/ml of stock solution. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare duplicate or triplicate standard points by serially diluting the Thrombin standard stock (20 ng/ml) 1:2 with equal volume of EIA Diluent to produce 10, 5, 2.5, 1.25, 0.625 and 0.313 ng/ml. EIA Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -200C and used within 30 days.


    I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns.

    Read More

    Question

    I received the abcam Thrombin Elisa Assay today. In anticipation of my experiment next week, I wanted to make the wash buffer solution. Instead of adding the 20X wash buffer concentrate to the dH2O, I added 10 mLs of the 10X diluent instead. I wanted to know what the diluent buffer components are so if I need more, I can make it.

    I appreciate your help.

    Read More
    Answer

    Thank you for contacting us.

    I heard back from the lab and they recommend that youdispose the Wash Buffer and the Diluent if you mixed them together.

    Wash Buffer is 20x and should be diluted with water (1 part Wash Buffer + 19 parts of water).
    Diluent is 10x and should be diluted with water (1 part of Diluent + 9 parts of water).
    Wewill send you 1bottle of the Wash Buffer and 1 bottle of the Diluent free of charge, but it may take us until next week to get these to you.
    I will be in touch again shorthly.

    Read More

    Question

    Is there any proteins in the storage buffer on the Thrombin standard that would run at about the same size as the thrombin protein itself?

    Read More
    Answer

    Thank you for calling Abcam earlier today.
    I have talked to the lab and they do not think there is protein the same size as thrombin in the buffer based, but we cannot be 100% sure. The exact components of the buffer are proprietary and so I am unable to provide that information.
    Please let me know if there is anything else I can help you with.

    Read More

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