antibodies into viable cells

David.C.McFarland@gsk.com David.C.McFarland at gsk.com
Wed Mar 27 20:24:20 EST 2002


I've come across several techniques lately that are supposed to be able to
get Abs into live cells but how can this be used for flow-based
intracellular staining?  More to the point, how to determine specificity?
Obviously, you aren't able to wash out the excess, do a blocking step, or
use an isotype control, right?  One reference I saw was using microscopy
to show localization/aggregation of the MAb in the cell, which is great
under the scope, but I just can't see how this could be a useful flow
technique.  Can someone enlighten me?

David McFarland
GlaxoSmithKline
----- Forwarded by David C McFarland/DEV/PHRD/SB_PLC on 27-Mar-2002 20:17
-----


"ray hester" <rhester at jaguar1.usouthal.edu>

26-Mar-2002 12:43




        To:     "Cytometry Mailing List"

        cc:
        Subject:        antibodies into viable cells



Dear group,

The following was sent to me directly so thought I would pass it along.

________________________________________________________
Hi Ray,
A colleague passed your question from the cytometry listserve on to me
and I thought I'd reply with a novel method I came across recently.
Feel free to post it.  In a recent Nature Biotechnology article (Morris
MC, Depollier J, Mery J, Heitz F, Divita G. A peptide carrier for the
delivery of biologically active proteins into mammalian cells. Nat
Biotechnol. 2001 Dec;19(12):1173-6.) they linked a couple of different
Abs to a Pep1 fragment, a protein transduction domain, and successfully
got them into cells.  High efficiency, low toxicity.
Regards,
Mark


C. Mark Lies, Ph.D.
Technical Marketing Scientist
Miltenyi Biotec, Inc.
12740 Earhart Ave.
Auburn, CA  95602

Toll free: (800)367-6227
Direct:    (530)887-5365
mlies at miltenyibiotec.com
www.miltenyibiotec.com




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