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

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.

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.

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.