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人Antithrombin III ELISA试剂盒(ab108801)

Reviews (1)Q&A (2)References (2)
ELISA: Antithrombin III Human ELISA Kit
  • ELISA: Antithrombin III Human ELISA Kit
  • ELISA: Antithrombin III Human ELISA Kit
  • ELISA: Antithrombin III Human ELISA Kit
  • ELISA: Antithrombin III Human ELISA Kit
  • Typical Standard Curve
  • Typical Standard Curve

Key features and details

  • Sensitivity: 0.71 ng/ml
  • Range: 6.3 ng/ml - 82.5 ng/ml
  • Sample type: Cerebral Spinal Fluid, Milk, Plasma, Saliva, Serum, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

概述

  • 产品名称

    人Antithrombin III ELISA试剂盒
    参阅全部 Antithrombin III/ATIII 试剂盒

  • 检测方法

    Colorimetric

  • 精确度

    批次内
    样品 n Mean SD CV%
    Overall 2.5%
    批次间
    样品 n Mean SD CV%
    Overall 9.4%

  • 样品类型

    Saliva, Milk, Urine, Serum, Plasma, Cerebral Spinal Fluid

  • 检测类型

    Sandwich (quantitative)

  • 灵敏度

    = 0.71 ng/ml

  • 范围

    6.3 ng/ml - 82.5 ng/ml

  • 回收率

    94 %

  • 检测时间

    4h 0m

  • 实验步骤

    Multiple steps standard assay

  • 种属反应性

    与反应: Human

  • 产品概述

    Abcam’s Antithrombin III Human in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Antithrombin III in urine, milk, saliva, cerebrospinal fluid, plasma, and serum.


    An antithrombin III specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently an antithrombin III specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Complex 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 antithrombin III captured in plate.


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

  • 经测试应用

    适用于: Sandwich ELISAmore details

  • 平台

    Microplate

性能

  • 存放说明

    Store at -20°C. Please refer to protocols.
  • 组件 1 x 96 tests
    100X Streptavidin-Peroxidase Conjugate 1 x 80µl
    10X Diluent N Concentrate 1 x 30ml
    20X Wash Buffer Concentrate 2 x 30ml
    40X Biotinylated Human Antithrombin III Antibody 1 x 150µl
    Antithrombin III Microplate (12 x 8 well strips) 1 unit
    Antithrombin III Standard 1 vial
    Chromogen Substrate 1 x 7ml
    Sealing Tapes 3 units
    Stop Solution 1 x 11ml

  • 研究领域

  • 功能

    Most important serine protease inhibitor in plasma that regulates the blood coagulation cascade. AT-III inhibits thrombin as well as factors IXa, Xa and XIa. Its inhibitory activity is greatly enhanced in the presence of heparin.

  • 组织特异性

    Found in plasma.

  • 疾病相关

    Defects in SERPINC1 are the cause of antithrombin III deficiency (AT3D) [MIM:613118]. AT3D is an important risk factor for hereditary thrombophilia, a hemostatic disorder characterized by a tendency to recurrent thrombosis. AT3D is classified into 4 types. Type I: characterized by a 50% decrease in antigenic and functional levels. Type II: has defects affecting the thrombin-binding domain. Type III: alteration of the heparin-binding domain. Plasma AT-III antigen levels are normal in type II and III. Type IV: consists of miscellaneous group of unclassifiable mutations.

  • 序列相似性

    Belongs to the serpin family.

  • 翻译后修饰

    Phosphorylation sites are present in the extracelllular medium.

  • 细胞定位

    Secreted > extracellular space.
  • Information by UniProt

  • 别名

    • ANT3_HUMAN
    • Antithrombin
    • Antithrombin III
    • Antithrombin-III
    • AntithrombinIII
    • AT 3
    • AT III
    • AT3
    • AT3D
    • ATIII
    • Heparin cofactor I
    • MGC22579
    • Serine (or cysteine) proteinase inhibitor clade C (antithrombin) member 1
    • Serine cysteine proteinase inhibitor clade C member 1
    • Serine proteinase inhibitor clade C member 1
    • Serpin C1
    • Serpin family C member 1
    • Serpin peptidase inhibitor clade C (antithrombin) member 1
    • SERPINC1
    • THPH7
    see all

  • 数据库链接

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab108801于以下的经测试应用

    “应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

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

    图片

    • ELISA: Antithrombin III Human ELISA Kit
      ELISA: Antithrombin III Human ELISA Kit

      Linearity: Antithrombin III in samples at different diutions, corrected for the factor of dilution (duplicates +/- SD).

    • ELISA: Antithrombin III Human ELISA Kit
      ELISA: Antithrombin III Human ELISA Kit

      Linearity: Antithrombin III in samples at different diutions, corrected for the factor of dilution (duplicates +/- SD), showing ng x mL-1 on y-axis.

    • ELISA: Antithrombin III Human ELISA Kit
      ELISA: Antithrombin III Human ELISA Kit

      Linearity: Antithrombin III in samples at different diutions, corrected for the factor of dilution (duplicates +/- SD).

    • ELISA: Antithrombin III Human ELISA Kit
      ELISA: Antithrombin III Human ELISA Kit

      Antithrombin III in cell culture supernatants (duplicates +/- SD), tested in the dilution range of 1:1-1:100.

    • ELISA: Antithrombin III Human ELISA Kit
      ELISA: Antithrombin III Human ELISA Kit

      Antithrombin III in human biological fluids (duplicates +/- SD). The samples were differently diluted (serum and Li-heparine plasma: 1e4-1e5; CSF: 1e2-1e3; milk: 10-100; saliva: 5-100; urine: neat).

    • Typical Standard Curve
      Typical Standard Curve

      Representative standard curve using ab108801.

    • Typical Standard Curve
      Typical Standard Curve

      Representative standard curve using ab108801.

    文献 (2)

    发表研究结果有使用 ab108801？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab108801 被引用在 2 文献中.

    • Huang M  et al. Identification of protein biomarkers in host cerebrospinal fluid for differential diagnosis of tuberculous meningitis and other meningitis. Front Neurol 13:886040 (2022). PubMed: 36003300
    • Cohen CT  et al. Human endothelial cells and fibroblasts express and produce the coagulation proteins necessary for thrombin generation. Sci Rep 11:21852 (2021). PubMed: 34750441

    客户评价及客户问答

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    1-3 of 3 Abreviews or Q&A

    Testing crossreactivity with rhesus serum and plasma

     Good Excellent 5/5 (Ease of Use)
    Abreviews
    An experience laboratory technician completed this Human Antithrombin III ELISA from Abcam, catalog number AB108801, lot #GR274235-1. The goal of the experiment was to analyze the linearity of dilutions of 2 different rhesus macaque EDTA-prepared plasma samples and determine the cross-reactivity of this kit for M. mulatta. Samples were stored prior to thaw at -80 degrees Celsius.

    Overall, the kit, like other Abcam ELISA kits, was easy to follow and simple to use.

    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 1:10 - 1:163840 (5 1:4 serial dilutions). At 1:2560, the subtracted OD’s were 0.1 or less likewise at 1:10 the OD was > than 1.0, putting it out of range of the assay (Charted blank-subtracted ODs can be found at the bottom of the corresponding image. Both serum and plasma were detected and in range of the assay @ 1:40, 1:160, and 1:640).

    We were disappointed in the low OD’s generated in the Standards. We determined on a more narrow window to find linearity in n=2 rhesus plasma for our second test. We also decided to use another TMB substrate instead of that provided in the kit to use a larger range of ODs (TMB High Sensitivity Substrate Solution, Cat# 421501, BioLegend).

    We diluted EDTA prepare plasma from n=2 Rhesus 1:100, 1:200, 1:400, 1:800, and 1:1600 in the second test. With the exception of the aforementioned TMB substrate, the rest of the protocol was performed as described in the kit’s instructions. The use of the higher sensitivity TMB certainly allowed a larger OD range, but contributed to large CVs at and below 6.25ng/mL. The samples’ subtracted OD values can be found in the corresponding image. A spike of 40ng/mL could not be detected in plasma that was diluted in these ranges (spiked after dilution to be in measureable range of standard).

    Based on these assays we can’t recommend the kit for detection of rhesus sample for 2 reasons. 1. We could not detect a linear range for the assay. However, since the higher sensitivity TMB substrate allowed for larger dilution ranges, a linear range may exist beyond 1:1600 if tested. 2. We could not detect spiked human AT3 in the presence of rhesus plasma.
    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

    提交于 Jul 01 2022

    Question

    Hi, I am trying to run the antithrombin kits (ab108801), but I cannot find the details of the quantity of antithrombin standard.
    Consequently, I cannot calculate how much diluent to add.
    Can you please email me the quantity of standard that is supplied, so that I can make this calculation and continue with my work?
    The product page just says "1 unit" is supplied. The protocol booklet makes no reference and just says to reconstitute to make a stock of 1.2 µg/ml.

    Read More
    Answer

    Please find below the information on how to prepare the standard for lots GR116626-1, Lot GR115289-1 and Lot GR106377-1 of the Antithrombin III Human ELISA Kit ab108801:


    · MIX Diluent Concentrate (10x): If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. Dilute the MIX Diluent Concentrate 1:10 with reagent grade water. Store for up to 1 month at 2 - 8ºC.

    · AT III Standard: Reconstitute the 4.8 ug of human AT III Standard with 4 ml of MIX Diluent to generate a stock solution of 1.2 ug/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. The stock solution (1.2 ug/ml) should be further diluted 1:6 with MIX Diluent to produce 200 ng/ml standard solution. Prepare duplicate or triplicate standard points by serially diluting the Standard solution (200 ng/ml) 1:2 with equal volume of MIX Diluent to produce 100, 50, 25, 12.5, 6.25, and 3.13 ng/ml. MIX Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -20ºC and used within the next 30 days.






    Standard Point
    Dilution
    [AT III] (ng/ml)

    P1
    1 part stock (1.2 ug/ml) + 5 parts MIX Diluent
    200.00

    P2
    1 part P1 + 1 part MIX Diluent
    100.00

    P3
    1 part P2 + 1 part MIX Diluent
    50.00

    P4
    1 part P3 + 1 part MIX Diluent
    25.00

    P5
    1 part P4 + 1 part MIX Diluent
    12.50

    P6
    1 part P5 + 1 part MIX Diluent
    6.25

    P7
    1 part P6 + 1 part MIX Diluent
    3.13

    P8
    MIX Diluent
    0.00









    Read More

    Question

    I am looking to quantify anti-thrombin in human urine samples and am potentially looking at 426 samples!

    Your Anti-thrombin III kit, ab108801 looks to be appropriate. How many plates would you recommend and what is the best quote you can give me? I haven’t had much luck with Abcam kits and technical help in the past and am hoping this will be an opportunity to restore my faith!

    Many thanks in advance,

    Read More
    Answer

    Thank for your email. I am sorry to hear that you have had bad experience with Abcam. Generally we are very flexible with our customers - our customer comes first than business so I am sure this time you will have the best technical support experience.

    The kit ab108801 is recommended to be used with 96 well plate. The experiment is set up in such a way that you would need 12 wells (6 in duplicate) for standard curve or 14 as given in table on page 9 and 2 wells for blank (duplicate). Now you are left with 80 wells.

    The 80 wells in one plate can be used for 40 samples in duplicate which is recommended or 80 samples if you are comfortable with pipetting etc.

    426 - 40 (in duplicate with one kit) = 386 samples

    If you are interested in duplicate then 40 samples are possible with first kit and 48 samples with each rest of the kits (96 wells to be used with each kit) in total 8. This means you would need 9 kits in total.

    My recommendation is to try the standard curve in at least one other kit as well to see the sample variations. In this case you would need total of 10 kits.

    Could you let me know if you are interested in buying 9-10 kits as recommended or have different opinion. I will then be happy to quote the kits with discount values.

    Thanks!

    Read More

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