Maleimide Quantification Assay Kit (Fluorometric) (ab112141)

Overview

  • Product nameMaleimide Quantification Assay Kit (Fluorometric)See all Maleimide kits ...
  • Tests
    1 x 200 test
  • Assay typeQuantitative
  • Product overview

    ab112141 uses a proprietary dye that has enhanced fluorescence upon reacting with a maleimide. The kit provides a sensitive, one-step fluorometric method to detect as little as 10 picomoles of maleimide in a 100 µL assay volume (100 nM; Figure 1). 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. Its signal can be easily read by a fluorescence microplate reader at Ex/Em = 490/520 nm. Compared to ab112140, this fluorometric assay is more sensitive, and has less interference from biological samples.

    Visit our FAQs page for tips and troubleshooting.

  • Notes

    ab112141 should be stored desiccated.

  • Tested applicationsFunctional Studiesmore details

Properties

    Applications

    Our Abpromise guarantee covers the use of ab112141 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    Functional Studies Use at an assay dependent concentration.

    Maleimide Quantification Assay Kit (Fluorometric) images

    • Standard curve: mean of duplicates (+/- SD) with background reads subtracted.
    • N-ethylmaleimide dose response was measured in a 96-well black plate with ab112141 using a microplate reader. As low as 0.1 µM (10 picomol/well) of maleimide can be detected with 10 minutes incubation time (n=3).

    Protocols

    References for Maleimide Quantification Assay Kit (Fluorometric) (ab112141)

    ab112141 has not yet been referenced specifically in any publications.

    Product Wall

    The quencher is still with the fluorophore after reacting with maleimide, thus the whole molecule is conjugated to the test sample. The sensing reaction only breaks the quencher moiety's conjugation system, but the quencher is still with the fluorophor...

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    I can confirm that the Maleimide Quantification Assay Kit (Fluorometric) (ab112141) can be used for measuring 6-8 samples at a time instead of 96. Please follow the instructions below:
    - Aliquot 500X MalemGreen DMSO stock solution (step 1), and st...

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    The lab confirmed that the MalemGreen component has a MW of 452.5 Da, and it is hydrophobic.

    Is this kit is specific to Maleimide?
    Yes

    Does this kit detect any other thiols?
    No

    How stable is this kit?
    Please use the fresh assay mixture within 1 hour. The 500X Maleimide Green Stock solution is stable at -20C.
    ...

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    Thank you for contacting us.

    The readings are really depending on the instruments. We use Molecular Devices's Gemini fluorescent plate reader, the background reading is around 1000-2000, the reading at 10 uM maleimide is ˜4000-7000.

    F...

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    Thank you for your reply.


    I have confirmed with the laboratory that regarding the reaction buffer, the pH of this cannot be altered.


    You are correct in that the first step is to prepare the reaction mixture and incubate ...

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    Thank you for contacting Abcam regarding ab112141.


    The reaction buffer, after adding it into the assay buffer has a pH of ˜7-7.5, so it shouldn't be a problem.


    I hope this information is helpful. Please do not hesitate t...

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    Thank you for contacting us. My colleague from the lab believes that you will be able to get a good signal by removing the solution, coverslipping your slides, and observing the signal under the microscope. This has not been tested, however, so I would...

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    Thank you for your reply.

    The pH for 2x assay mixture is ˜ 7.4.

    Please let me know if there is anything else I can help you with.

    Thank you for yourreply.

    The pH for the reaction buffer (component B) is very basic (˜ 12), and the pH for the assay buffer (component C) is 7.

    Please let me know if there is anything else I can help you with.

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