Question (67023) | Histone H3 (K4) Methyltransferase Activity Quantification Assay Kit (ab113452)

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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

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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