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Histone H10 Human, Recombinant > FAQ |  | Histone H10 Human, Recombinant FAQ
See the Epigenetics FAQ also.
Q1: Do the histones need to be reconstituted?
Q2: What are the recommended histone storage conditions?
Q3: Are the histones fusion proteins or tagged proteins?
Q4: Can the histones be used as substrates for protein modification enzymes? Which ones?
Q1: Do the histones need to be reconstituted?
A1: No. The histones are sold in solution at 1mg per ml in 20mM NaPO4 (pH 7.0), 0.3M NaCl, 1mM EDTA, 1mM DTT.
Q2: What are the recommended histone storage conditions?
A2: We recommend storing the histones at -20°C. The histone solution will freeze because there is no glycerol in the storage buffer. Repeated freezes and thaws have not been an issue in our hands.
Q3: Are the histones fusion proteins or tagged proteins?
A3: The histones do not have any tag and are not produced or purified as a fusion protein. They are purified from an E. coli strain that carries a plasmid encoding the cloned human histone gene. Each histone’s protein sequence and genbank accession number can be found on its NEB product webpage and on its datacard.
Q4: Can the histones be used as substrates for protein modification enzymes? Which ones?
A4: They would presumably serve as excellent substrates for any histone modification enzyme. We ensure that the recombinant histones are not modified at any potential modification site by producing them in E.coli and then further checking each histone’s mass by ESI-TOF MS (Electrospray Ionization-Time of Flight Mass Spectrometry).
The histone modification enzymes that we have tested are:
Histone H1°: CDK2-cyclin A protein kinase (NEB #P6025S) and Cdc2-cyclin B protein kinase, also known as cyclin-dependent kinase 1 (CDK1) or M-phase promoting factor (MPF) (NEB #P6020S)
Histone H3.1, H3.2 and H3.3: G9a Methyltransferase (NEB #M0235) and SET7 Methyltransferase (NEB #M0233)
Histone H4: PRMT1 Methyltransferase (NEB #M0234)
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