You can also use a product called Nycoprep Mixer from Nycodenz which is just
mixed 1:1 with the blood, mixed and spun at high speed. Similarly neutrophils
are in the pellet and you have to lyse the red blood cells.
Good luck
Gerhard
In article <36C4402E.B3CF2F58 at slu.edu>,
"Michael J. Richards" <richarmj at slu.edu> wrote:
> I have used this gradient some years ago for chemotaxis studies and obtained
> very pure neutrophil preps. As I recall you first dilute the anticoagulated
> whole blood 50/50 with 150mM saline and overlay your ficoll-hypaque gradient
and
> after spin collect your bottom pellet which is neutrophil/RBC mix and then
lyse
> off the RBC's
>> Jeffrey & Sandy Marion wrote:
>> > Eric Park wrote:
> > >
> > > I'm encountering some problems when I try to isolate neutrophils from my
> > > blood. I'm using a Ficoll-Hypaque solution when I centrifuge the blood.
> > > This technique works great for several other people but not for me. A lot
> > > of RBCs are remaining in the neutrophil layer and not settling down to the
> > > bottom of the tube. Does anyone know why this is happening to my blood?
> > >
> > > Personal info:
> > > Blood type is B+ Rh+
> > > Asian male 22 years old
> > >
> > > Insights would be helpful since this is a mildly annoying problem in my
> > > research by begging for volunteers instead of tapping my own blood.
> >
> > In the world of flow cytometry, many technologists utilize ficol
> > histopaque medium to recover lymphocytes from anticoagulated whole blood
> > or lymphocytes from lymph nodes. Spun at 1500 rpms for 30 minutes
> > yields an excellent recovery of these T and B cells with very little
> > monocyte or other leukocyte contamination.
>> --
> Michael J. Richards
> Research Biochemist
>>
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