extraction of histones

Alexander N. Kukushkin kan at mmcd.cyt.ras.spb.ru
Thu Jul 31 11:13:59 EST 1997


>   I am using the Dignam method (NAR, 1983) for obtaining cytoplasmic/nuclear
>   extracts.
>   The hypertonic buffer employed for nuclear extraction has 0.5 M NaCl. I
>   think that in such conditions I recover histone H1, but I do not known
>   what happens with core histones. I should be grateful to receive
>   obvservations about it.
>
>   Thanks in advance
>
>   Anxo Vidal
>   Dept. Physiology
>   School of Medicine
>   University of Santiago de Compostela
>   SPAIN
>
Many years ago I have carried out selective extration of lysine-rich
histone H1 from the nuclei by a solution containing 0.3-0.35 M NaCl, pH3.0
according to Villeponteaux B. et al, Biochemistry, 1978, 17, 5532. If the
extraction was performed by solution with 0.4 M NaCl and higher the core
histones released partially and nucleosome structure of chromatin was
disturbed. Thus, I think that nuclear extracts with 0.5 M NaCl are to
contain many core histones.
Alex



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