Help! CD3+ Flow Cytometry

Jorg Kirberg kirberg at nki.nl
Fri Aug 9 10:18:59 EST 1996


In article <4uec2f$6pj at newsbf02.news.aol.com>, snowtrek at aol.com (SNOWTREK)
wrote:

> Greetings, Colleagues: Nothing too exotic about this...my chore is the
> binding of a humanized  anti-CD3 monoclonal antibody to a CD3+ cell,
> followed by an anti-human kappa FITC conjugate. Seems simple enough, but
> the CD3 positive cells I've used - HUT78, Jurkat - look like hell, with
> broad gnarly histograms. What I need is a cell that's 1) high in CD3
> density (will yield high median fluorescence intensity), will give me 2)
> nice narrow histograms with low CVs on the FL1 parameter, and 3) is
> readily available, ATCC best. Any suggestions? Anybody ever use Daudi's? I
> think HPB-ALLs might work well, but I'm unaware of a commercial source.
> Can't use HUT102s, possible HTLV-1 +. Muchas gracias for any suggestions.
> - Bob/snowtrek at aol.com
Dear Bob,

sorry, but why don't you simply get a bit of blood from someone in the lab
and use those PBLs ? (in a two colour FACS analysis to differentiate the
kappa positive B cells).

jorg


kirberg at nki.nl



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