Need method for isolating pure T from Human PBLs

Derek V. Chan chan4 at fas.harvard.edu
Fri Apr 7 20:24:35 EST 1995


In <3m3vuj$prl at netnews.upenn.edu>, David Peritt <Peritt_d at a1.mscf.upenn.edu> writes:
>In article <3m2io4$b3a at decaxp.harvard.edu> Derek V. Chan,
>chan4 at fas.harvard.edu writes:
>>Anybody know of a good, easy way to purify T-cells from PBLs?  We want 
>>to remove NK cells and any other cells that may lead to nonspecific
>lysis of
>>target cells in a CTL assay.  We've utilitzed Percol T gradient
>separation;
>>however, it appears that the population we have isolated is still not
>totally
>>pure T....
>
>
>I assume you have pondered the use of the classical techniques such as
>negative or poistive panning, complement killing and FACS/MACS sorting.


Yep!  Unfortunately, panning only yields at best 97% clean....

C' killing doesn't deplete out everything.

We want to get rid of the LAK precursor population.  I'm considering anti-CD56
C' depletion or magnetiic bead separation or a column...


Derek V. Chan
chan4 at fas.harvard.edu
dvchan at netcom.com




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