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Assays for histone methylation

Related

  • Products for your research
    • ChIP grade antibodies
      • ChIP & ChIPseq kits
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          • Extraction kits
            • Methylated ChIP kits
              • HDAC & SIRT activity kits
                • Histone resources
                  • Histone modifications
                    • Methods to study histones
                      • Advantages of our anti-H3
                        • Most cited H3 antibody

                          Solutions for measuring histone methyltransferase and demethylase activity

                          Histone methyltransferase activity assays

                          Assay development for histone methyltransferases (HMTs) is difficult because they have often have low kcat (turnover is typically below 1 min-1) and low KM values for their methyl donor S-adenosylmethionine (SAM).  HMT assays therefore have need to have very high sensitivity. Many HMT activity assays measure S-adenosylhomocysteine (SAH) formation, a by-product of the methylation reaction. This means that they are not specific for any HMT, and can only be used with purified protein/protein complexes. They also often use 3H-SAM as a donor, and so involve handling radioactive material.

                          Our HMT activity assays overcome these difficulties by using antibodies to detect the methylated product, giving you: 

                          • Easy quantitative detection, without the need for radioactivity 
                          • The ability to use nuclear extracts as well as purified proteins (as the assay is specific for the modification of interest) 
                          • Microplate format with simple colorimetric or fluorometric readouts 
                          • Data in 3 hours ​​​​


                          Activity measuredProduct nameProduct code
                          Methylation of histone H3 on K4Histone H3 (K4) Methyltransferase Activity Quantification Assay Kitab113452
                          Methylation of histone H3 on K9Histone H3 (K9) Methyltransferase Activity Quantification Assay Kitab113453
                          Methylation of histone H3 on K27Histone H3 (K27) Methyltransferase Activity Quantification Assay Kitab113454

                          Histone demethylase activity assays

                          Histone demethylase assays usually measure the formation of formaldehyde, a by-product of demethylation. They are therefore susceptible to interference from detergents, thiol groups and a range of ions. In addition, these assays are not specific for any demethylase and can only be run with purified protein.

                          • Our histone demethylase assays circumvent these issues by directly measuring the formation of demethylated product, giving you:​
                          • Increased sensitivity (20–1000 fold) over formaldehyde-based assays 
                          • More accurate data – no interference from thiols, detergents or ions
                          • Compatibility with nuclear extracts as well as purified protein (due to the assay’s specificity for the modification of interest)
                          • Microplate format with simple colorimetric or fluorometric readouts 
                          • 3 hour assay time


                          Activity measured


                          Product name
                          Product code
                          Demethylation of histone H3K4me2Histone Demethylase (H3K4) Activity Quantification Assay Kitab113455
                          Demethylation of histone H3K4me2 and H3K4me3 by KDM1 / LSD1

                          KDM1/LSD1 Activity Quantification Assay Kit (Colorimetric) 

                          KDM1/LSD1 Activity Quantification Assay Kit (Fluorometric)

                          ab113459

                          ​                                          

                          ab113460

                          Demethylation of H3K9me3 by:
                          KDM4A/JHDM3A/JMJD2A, KDM4B/JMJD2B, KDM4C/GASC1/JMJD2C and KMD4D/JMJD2D

                          KDM4/JMJD2 Activity Quantification Assay Kit (Colorimetric)

                          KDM4/JMJD2 Activity Quantification Assay Kit (Fluorometric)

                          ab113461


                                ab113462

                          Demethylation of H3K4me2 and H3K4me3 by KDM5/JARID

                          KDM5/JARID Activity Quantification Assay Kit (Colorimetric) 

                          KDM5/JARID Activity Quantification Assay Kit (Fluorometric)

                          ab113463

                                 ​

                                          ab113464

                          Demethylation of H3K27me3 by KDM6A/UTX and KDM6B/JMJD3

                          KDM6A/KDM6B Activity Quantification Assay Kit (Colorimetric) 

                          KDM6A/KDM6B Activity Quantification Assay Kit (Fluorometric)

                          ab156910

                          ​

                                 ​ab156911​


                          View our full list of activity assays for writers and erasers of the histone code.

                          If you need to measure the total amount of  histone or histone modification in your sample, try our quantification kits. View the complete list here.

                          ​

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