On Fri, 12 Feb 1999 08:47:53 -0600, "Michael J. Richards"
<richarmj at slu.edu> wrote:
>For PBMC I have used just the standard ficoll/hypaque type gradient on whole
>blood diluted 50/50 with 150mM saline. This can then be followed by using a
>percoll(pharmacia) type gradient.
>>Conny Stuelten wrote:
>>> I am trying to isolate monocytes from human PBMC with density gradients. In
>> the beginning I have used OptiPrep-gradient and obtained a
>> purification from 60%. Now I utilize an Iopromid-gradient (Biochrom) with
>> the density 1,068 g/l, but I got no results (no seperation). Has anyone
>> worked with Iopromid-gradient or with another Monocyte-gradient?
>>>> Thanks Conny
You need to follow this up with overnight (or even shorter) adherence
to plastic, followed by washes to remove nonadherent cells.
Otherwise, as far as I know you can't exclude the presence of other
lymphocyte types.
As I recall, you can go straight to plastic adherence from
ficoll/hypaque without other fancy gradient media and get good preps,
although I do know people that have used specialty gradients with good
success (will find out for you).
Terry L. Davis
