need protocol to isolate nuclei from lymphocytes

Ken Frauwirth BioKen frauwirt at notmendel.Berkeley.EDU
Thu Oct 13 21:36:11 EST 1994


In article <mrtour-1310941204460001 at mac-133.k-g2.ski.mskcc.org>,
michelle tourigny <mrtour at stud.med.cornell.edu> wrote:
>I have been trying to isolate nuclei from lymph or spleen for a DNase 1
>hypersensivity assay and with either 0.5% or 1% NP-40 incubation at 4
>degrees C I have been unable to have consistent lysing or nuclei.  Any
>suggestions?
>
>Michelle Tourigny, grad student
>
>-- 
>Michelle Tourigny
>mrtour at stud.med.cornell.edu
>
>Elmo says:  "But I'm not a scary monster"

You may want to try disrupting the cells with a Dounce Homogenizer in a
hypotonic solution.  This is how I always used to prepare nuclear extracts
(although I was not using lymphocytes).  This is a pretty standard procedure,
and there is a protocol in Current Protocols in Molecular Biology in the 
section on preparing nuclear extracts.

BioKen
-- 
Ken Frauwirth (MiSTie #33025)       _           _
frauwirt at mendel.berkeley.edu       |_) *    |/ (_ |\ |
Dept. of Molec. & Cell Bio.        |_) | () |\ (_ | \|  
Univ. of Cal., Berkeley          I had a .sig, but I broke it.



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