Thiol Quantification Assay试剂盒(Fluorometric) (ab112158)

概述

  • 产品名称Thiol Quantification Assay试剂盒(Fluorometric)
  • 测试
    1 x 200 test
  • 样品类型
    Urine, Plasma, Cell culture extracts
  • 检测类型Quantitative
  • 产品概述

    The detection and measurement of free thiol (such as free cysteine, glutathione, and cysteine residues) is one of the essential tasks for investigating biological processes and events in many biological systems. There are a few reagents or assay kits available for quantitating thiol content in biological systems. All the commercial kits either lack sensitivity or have tedious protocols.


    ab112158 provides an ultrasensitive fluorometric assay to quantitate thiol content that exists in a small molecule. The proprietary non-fluorescent dye used in the kit becomes strongly fluorescent upon reacting with thiol.
    ab112158 can detect as little as 1 picomole of cysteine or GSH in a 100 µL assay volume (10 nM). The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. The thiol sensor used in the kit generates a strongly fluorescent adduct upon reacting with a thiol compound. This adduct has spectral properties almost identical to those of fluorescein. In addition, both absorption and emission spectra of the thiol adduct are pH-independent, making ab112158 highly robust. The signal can be easily read by a fluorescence microplate reader at Ex/Em = 490/520 nm.


     


    Please note it is not possible to do a quantification in comparison to the standard.

    Visit our FAQs page for tips and troubleshooting.

  • 说明

    ab112158 should be stored desiccated.

  • 经测试应用适用于: Functional Studiesmore details

性能

    应用

    Our Abpromise guarantee covers the use of ab112158 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
    Functional Studies Use at an assay dependent concentration.

    Thiol Quantification Assay Kit (Fluorometric) 图像

    • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

    • GSH measured in cell lysates showing quantity (nmol) per 1 mln cells.

      Samples with the concentration of 1e8 cells/mL were used. Samples were diluted 100-1000 fold.

    • GSH measured in biological fluids. Samples were diluted 10-1000 fold.

    • GSH and Cysteine dose responses were measured in a 96-well black plate with ab112158 using a microplate reader. As low as 10 nM (1 pmol/well) of GSH or Cysteine can be detected with 10 minutes incubation time (n=3). The insert shows the low levels of thiol detection.

    实验方案

    Thiol Quantification Assay Kit (Fluorometric) (ab112158)参考文献

    ab112158 has not yet been referenced specifically in any publications.

    Product Wall

    1. The kit can only quantify thiol groups, not disulfide bonds.

    2. DTT cannot be used as reducing agent because it contains -SH groups, and these will interfere with this assay. That is also the case for beta mercaptoethanol. If either has t...

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    This kit is very sensitive to presence of DTT, beta mercaptoethanol or guanidine HCl. Even nanomolar quantities of these reagents will negatively effect the results. Please ensure your samples are free of these reagents before using this kit.


    The Thiol Quantification Assay Kit (Fluorometric) (ab112158) should be compatible with both Urea or Guanidinium-HCl and should not interfere with the assay.

    The kit is provided with glutathione (GSH) as a standard. This molecule contains ...

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    Thank you for your email and your interest in our products.

    I can confirm that regrettably TCEP and all other reducing agents such as DTTor b-mercaptoethanol will interfere with the Thiol Quantification Assay Kit (Fluorometric) (ab112158).
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    Thank you for contacting us.

    I have heard back from the lab with the following advice:

    A solid black microplate (black walls and black bottom) would bepreferably recommendedover a black-walled microplate with this assay.
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    Abreviews
    the kit is very easy to use. it contains very low amount of assays though.
    Username

    Mrs. smadar tabenkin

    Verified customer

    提交于 Sep 03 2014

    I can confirm that using bacterial samples should be ok as long as the bacteria is lysed. The detection limit of the kit is 10 nM, so as long as the there is 10 nM cysteine differences among the mutations, it should be OK. Please note this has not bee...

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    Thank you for contacting us.

    Yes, the kit can be used for endpoint read. I recommend you read at 15min, 30 min and 1 hour see which one is best for your experiment. The Ex/Em=490/520 nm.


    I hope this information is helpful to you...

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    Please see my math below calculating the minimum volume of your sample needed if we assume the final molar concentration of free cysteine is 25% of the molar concentration of cysteine originally in your sample (50% from 1:1 reduction in DTT or TSEP, th...

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    Theoretically TCEP should not effect this assay but it does react with the dye. TCEP seems to react with a lot of the things but unfortunately we are not sure how this occurs.

    1-10 of 17 Abreviews or Q&A

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