EpiSeeker DNMT Activity Quantification Kit (Colorimetric) (ab113467)

Overview

  • Product nameEpiSeeker DNMT Activity Quantification Kit (Colorimetric)See all DNMT kits ...
  • Assay typeQuantitative
  • Assay time
    3h 45m
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Plants
    Predicted to work with: all Mammals, Fungi
  • Product overview

    DNMT (DNA methylases) is a family of enzymes responsible for the addition of methyl groups on the cytosine ring at the 5' position of a CpG dinucleotide. The DNMT family is required for the establishment and maintenance of DNA methylation patterns.

    EpiSeeker DNMT Activity Quantification Kit (Colorimetric) (ab113467) allows the user to colorimetrically measure DNA methyltransferase activity or inhibition at extremely fast speed on a 96-stripwell microplate. The kit is ready-to-use and provides all the essential components needed to carry out a successful DNMT activity/inhibition experiment without the need for radioactivity or any special equipment.

    Not sure if this is the right product for you? Check out our EpiSeeker DNMT Product Guide for help.

  • Notes

    Store EDM3, EDM4, EDM6, and EDM7 at –20°C away from light.

    Store EDM1, EDM5, EDM8, and the 8-Well Assay Strips at 4°C away from light.

    Store all remaining components (EDM2, EDM9, and the Adhesive Covering Film) at room temperature away from light.

    All components of the kit are stable for 6 months from the date of shipment, when stored properly.

  • Tested applicationsFunctional Studiesmore details

Properties

    Applications

    Our Abpromise guarantee covers the use of ab113467 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.

    EpiSeeker DNMT Activity Quantification Kit (Colorimetric) images

    • Demonstration of high sensitivity and specificity of the DNMT activity assay achieved by using nuclear extracts with ab113467. Nuclear extracts were prepared from MCF-7 cells using ab113474.
    • Demonstration of high sensitivity and specificity of the DNMT activity/inhibition assay achieved by using recombinant DNMT1 with ab113467.

    Protocols

    References for EpiSeeker DNMT Activity Quantification Kit (Colorimetric) (ab113467)

    ab113467 has not yet been referenced specifically in any publications.

    Product Wall

    DNMT ACTIVITY CUANTIFICATION KIT
    PROTOCOL for ADIPOSE TISSUE PROTEINS
    • EDM1, EDM2, EDM3 were prepared following manual instructions.
    • The amount of protein obtained in the nuclear extraction of this kind of tissue was very low. Therefore, we decided to use 20 g of nuclear extract and complete with a total volume of 100 L/well. (After different probes we saw that we need this amount of protein, and we safe the proportion, 1-5 L of nuclear extract and 40-45 L of EDM3, completing with 100 L/well.)
    • For blank and positive control wells, the volume was changed to 100 L/well.
    • The strip-well microplate was covered with the adhesive film and was incubated in an oven at 37ºC with continued shaking during 3 hours.
    • The washes were done following the instructions in all steps.
    • EDM5 were prepared in a dilution rate of 1:500 and incubated in the same conditions that in the previous step (oven at 37ºC with continued shaking during 1 hour).
    • EDM6 were prepared in a dilution rate of 1:1000 and incubated in the same conditions that in the previous step (oven at 37ºC with continued shaking during 30minutes).
    • EDM7 were prepared in a dilution rate of 1:2000 and incubated in the same conditions that in the previous step (oven at 37ºC with continued shaking during 30minutes).
    • 100 L/well of EDM8 were added and the microplate was incubating at room temperature away from direct light about 30 minutes.
    • EDM9 were added to stop the enzyme reaction.
    • Activity calculation was done following the instructions.
    Username

    Miss. Ana Jadraque

    Verified customer

    Submitted Jun 25 2014

    DNMT activity in mouse brain

    Good Good 4/5 (Ease of Use)
    Abreviews
    The kit is easy to use and contains all components needed to perform the colorimetric ELISA. Mouse brain samples were used with this kit. Nuclear extract was prepared (with a non abcam kit) from the previous collected mouse brain samples. From each sample 15 µg of nuclear extract was used to perform the ELISA. From here on the abcam protocol was followed. The blank an positive controls were included, and the positive control had the highest signal in my study design. Between different individual animals there was a low variation, speaking for the high sensivity and reproducibility of the kit.
    Username

    Abcam user community

    Verified customer

    Submitted Jun 07 2013

    Thank you for your message.

    I am sorry that the vial you received with this kit did not contain the full amount of the product and I apologize for the inconvenience. I have issued a free of charge replacement vial with the order number of ##...

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    Thank you for your reply.

    As far as I am aware there isn’t too much difference between coating the ELISA plates with proteins or DNA, but I did find the following publication that I think will be useful to you:

    http://onlineli...

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    Thank you for contacting Abcam.

    I have talked to the lab about the questions you had for ab113467. Please find the answers to these below:

    1 - Is the microplate flat bottom or round? It is the flat bottom.

    2 - What coating...

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    Thank you for contacting Abcam and you rpatience whilst I contacted the lab about your enquiry.

    The Dnmt substrate is a synthesized double stranded DNA polynucleotide ( >300 bps)containing >25%CpGs.



    If there is anyt...

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    Thank you for contacting Abcam.

    All of the information that we have about this kit is available on the webpage’s for the product. Currently there are no know references for this kit, but we are happy to assure you that it is covered unde...

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