Overview

  • Product name
    Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit
    See all Histone Methyltransferase kits
  • 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 9 (K9) mediates heterochromatin formation by forming a binding site for HP1 and also participates in silencing expression at euchromatic sites. In mammalian cells, histone lysine methyltransferases such as SuVar39h and ESET among others are responsible for methylation of lysine 9 at histone H3.

    Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit (ab113453) allows the user to measure the activity or inhibition of individual histone methyltransferase (HMT) that specifically target histone H3 at lysine 9 (H3K9). The kit is ready-to-use and provides all the essential components needed to carry out a successful HMT activity/inhibition quantification experiment without the need for radioactivity or any special equipment.

    ab113453 recognizes methylated H3 at lysine site 9.

Properties

Images

  • Nuclear extracts were prepared from MCF-7 cells using the ab113474 and H3-K9 specific histone methyltrasferase activity (G9a) was measured. Purified G9a was spiked into the nuclear extract that has low concentration of nuclear proteins by dilution.

Protocols

References

ab113453 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us.
Here are the answers to your questions:
1. Peptide.
2. Yes, avidin-coated.
3. This information cannot be disclosed currently.
4. No. unless a purified enzyme or extract with the over-expressed specific enzyme are used.
5. Yes.
6. The standard is methylated H3K9 peptide
7. G9a
8. salt buffer with protein stabilizers
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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A standard curve was plotted by plotting the blank corrected OD450 for three different HMT standard concentrations. This kit works well and is specific for H3K9 Me-transferase.

Abcam user community

Verified customer

Submitted Mar 10 2017

Answer

Thank you for contacting us.
Here are the answers to your questions:
1. After dilution, each diluted component should be used within the same day.
2. At 4C for same day.
3. for longer time storage, as it will not be frozen at -20C.
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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Answer

Thank you for contacting us.
Here are the answers to your questions:
1. 100,000-500,000 cells or 10-50 mg tissues.
2. It is possible but results are not guaranteed.
3. It can be stable for 30 min after adding stop solution.
4. It may not work at 470 nm.
5. Enzyme or nuclear extract reaction step should be finished within 1-3 h at 37C. Capture antibody step could be overnight at 4C. However the background may be increased.
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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Answer

Thank you for contacting us and thank you for your patience.

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

1. Yes, this kit is suitable for the conditions mentioned (rat oligodendrocytes with and without inhibitors for MT) if the sufficient amount of nuclear protein can be obtained from rat oligodendrocytes andis used for each assay point ( 3-5 ug). Rat samples have however not yet been tested, but should work theoretically.

2. Purified G9a was spiked into the nuclear extract that has low concentration of nuclear proteins by dilution - for obtaining the image.

3. Unfortunately, a testing discount is not possible at the moment for this kind of kits. We offer them for antibodies and proteins as well as ELISA kits. I'm sorry about this.

4. The antibodies are specific to methylated H3 at lysine site 9.

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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Answer

Thanks for your inquiry. Yes, the kit ab113453 is suitable for measuring H3K9 activity from various species including Drosophila based on the working principle and antibody's specificity. Please let me know if you require further assistance.

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

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