Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit (ab113452)

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Overview

  • Product name
    Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit
    See all Histone Methyltransferase kits
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay type
    Quantitative
  • Assay time
    3h 00m
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Mammals
  • Product overview

    Methylation of histone H3 at lysine 4 (K4) is thought to be a global epigenetic mark in euchromatin and a mediator of activated transcription. In mammalian cells, several SET-domain containing histone lysine methyltransferases such as MLL1 are responsible for adding one, two or three methyl groups at lysine 4 of histone H3.
    Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit (ab113452) allows the user to measure the activity or inhibition of individual histone methyltransferases that specifically target histone H3 at lysine 4 (H3K4). The kit is ready-to-use and provides all the essential components needed to carry out a successful histone methyltransferase activity/inhibition quantification assay without the need for radioactivity or any special equipment.

  • Platform
    Microplate reader

Properties

Images

  • Functional Studies - Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit (ab113452)
    Functional Studies - Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit (ab113452)
    Nuclear extracts were prepared from MCF-7 cells using the ab113474 and H3-K4 specific histone methyltrasferase activity (SET7/9) was measured.

Protocols

References

ab113452 has not yet been referenced specifically in any publications.

Publishing research using ab113452? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Question

Dear Sir/Madam, Thank you for your quick reply and clarification. I would like to ask for other techical information: 1. What is the usefull concentration range of purified HMT enzyme (H3-K4 specific) for this kit (ab113452)? 2. Can I use the product ab80377 "SMYD3 protein (Active)" as purified recombinant protein in this kit? Thank you Your Sincerely

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Answer

Thank you contacting us with these further questions. I have spoken with the lab about a usefull concentration for the purified HMT enzyme. Based on tests, we conclude that the useful concentration range can be from 10 ng to 200 ng/well (30 ul). I also wanted to let you know that SMYD3 (ab80377), as di-and tri-methyltransferase against H3-K4, can be definitely used with this kit. If you have any other questions or there is anything else we can do to to help you reach your research goals, please contact us.

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Question

Dear Sir/Madam, I would like to have some techical information about your product ab113452 EpiSeeker Histone Methyltransferase H3(K4) Activity Quantification Assay Kit. - Can I use a purified HMT enzyme rather than nuclear extract to carry out my assay? - If not. Can you suggest an other colorimetric assay that allows to do

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Answer

Thank you for contacting us. I have had a chance to speak with the lab regarding your question. They have told me that the purified HMT enzyme (H3-K4 specific) can be used in this assay kit. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

Dear Sir/Madam,
I would like to have some technical information about your product
ab113452 EpiSeeker Histone Methyltransferase H3(K4) Activity
Quantification Assay Kit.
1. Could you indicate what is biotinylated substrate (EHM4) used for?
2. For each point (2ul) it need to obtain a different concentration of
HMT standard (ng/ul).
Could you indicate how to do standard curve (with an example)?
Looking forward to hearing you.
Thank you in advance
Yours sincerely

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Answer

Thank you for your enquiry regarding ab113452.
1. The biotinylated substrate is unmethylated histone peptide and used for H3-K9 specific histone methyltransferase reaction.
2. For generating a standard curve, first, dilute HK5 HMT Standard with HK2 Assay Buffer to 5 ng/µl by adding 5 µl of HK5 to 5 µl of HK2. Then, further prepare five concentrations by combining the 5 ng/µl Diluted HK5 with HK2 into final concentrations of 0.2, 0.5, 1.0, 2.0, and 5.0 ng/µl according to the following dilution chart. Then add 2 ul of diluted HK5 at different concentrations to each assay well.
Tube HK5(5 ng/µl) HK2 Resulting HK5 Concentration
1 0.5 µl 24.5 µl 0.1 ng/µ
2 1.0 µl 24.0 µl 0.2 ng/µl
3 1.0 µl 9.0 µl 0.5 ng/µl
4 1.0 µl 4.0 µl 1.0 ng/µl
5 2.0 µl 3.0 µl 2.0 ng/µl
6 4.0 µl 0.0 µl 5.0 ng/µl
I hope this will be useful for you.

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