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Maleimide Quantification Assay Kit (Fluorometric) (ab112141)

Q&A (14)

Overview

  • Product nameMaleimide Quantification Assay Kit (Fluorometric)See all Maleimide kits ...
  • Tests
    1 x 200 test
  • Sample type
    Cell culture extracts, Adherent cells, Suspension cells
  • 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 Studies more 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 Notes
Functional Studies FuncS

Maleimide Quantification Assay Kit (Fluorometric) images

  • Functional Studies - Maleimide Quantification Assay Kit (Fluorometric) (ab112141)
    Functional Studies - Maleimide Quantification Assay Kit (Fluorometric) (ab112141)
    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.

Publishing research using ab112141 ? Please let us know so that we can cite the reference in this datasheet

Product Wall

Submit an Abreview for ab112141 Submit a Question for ab112141

Displaying 1 - 10 of 14 results for Abreviews and Q&A

10
Votes: Highest First

What is the size of the MalemGreen indicator? Is it hydrophilic?


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

Customer is wondering if this kit is specific to Maleimide or if it detects any other thiols? How stable is this kit? Can it survive centrifuge filtering?

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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i am getting background of 10000 fluoresence units staying within the 30 min incubation, is this too high?

for data analysis, am I supposed to be taking the log of both the x and y axis to plot the data points?

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.

For ...

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Here is some raw data:









STANDARDS



Undil



1/5 Diluted







1



2



3 ...

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Thank you for your patience.

I have got a reply from the lab with some tips on the optimisation.

From the data, The standard is very good, the only sample works is column 3E & F. All other samples have lower counts. This is most li...

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Thank you for the reply...The reaction buffer is to activate the sensor, so after the sensor gets activated, can i lower the pH by adding another buffer to about 7. Based on the protocol you have for this product, first is to prepare the reaction mixture ...

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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 for...

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Based on the Q&A you provide in your website regarding this kit, the pH of the reaction buffer is pretty basic. Because my polymer can't dissolve in that high pH, i am afraid that my polymer will just precipitate out once i added the reaction buffer to my...

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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 to c...

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I would like to use this kit for detection of maleimide that has been coated on the surface of a matrix. Does this kit generate a fluorophore that is soluble or that is affixed to the maleimide?

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 re...

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As I just discussed with you on the phone, we are trying to estimate the amount of maleimide linker installed on a negatively charged polymer.
Theoretically, there should be more than 100 micro molar maleimide linker (6.5% in molar ratio against the ...

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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.

As I just discussed with you on the phone, we are trying to estimate the amount of maleimide linker installed on a negatively charged polymer.
Theoretically, there should be more than 100 micro molar maleimide linker (6.5% in molar ratio against the ...

Read More

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.

As I just discussed with you on the phone, we are trying to estimate the amount of maleimide linker installed on a negatively charged polymer.
Theoretically, there should be more than 100 micro molar maleimide linker (6.5% in molar ratio against the ...

Read More

Thank you for sending your data yesterday.

I passed your data along to the lab so that they could try and help understand what was going on. From the information that you sent, I believe that you samples were a Maleimide-PEG linker attached to a...

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