getting antibodies into viable cells

Laird Bloom LBloom at Phylos.com
Fri Mar 22 09:23:21 EST 2002


We have tried a lipid reagent called BioPorter (from Gene Therapy Systems)
and a peptide-based reagent called Chariot (from Active Motif).   Chariot
was not reliable in our hands.  BioPorter got antibodies into HeLa cells
nicely, but they (Alexa 594 conjugated anti-IgG Ab)  tended to be localized
in punctate spots around the nucleus. We didn't look at antibodies that are
expected to localize to specific places, but another Alexa 594-conjugated
protein that we tried should have been localized to the periphery of the
cell and still showed the punctate perinuclear staining.  We saw clear
evidence of its expected function in the periphery, however, suggesting that
there is a fraction of the protein that gets to the right place despite the
unexpected localization of the bulk of the protein.  The data wouldn't
convince a skeptic, though.

Laird Bloom

Phylos, Inc.
128 Spring St.
Lexington, MA  02421
tel. (781) 862-6400 ext. 253
fax (781) 402-8813
www.phylos.com



> ----------
> From:		ray hester
> Sent:		Thursday, March 21, 2002 9:20 AM
> To:	Cytometry Mailing List
> Subject:	getting antibodies into viable cells
>
>
> Greetings,
>
> A recent discussion on the Confocal Listserv, concerning DAPI staining of
> viable cells, prompts me to send the following from a colleague:
>
> Is there a way to deliver antibodies, or their fragments, into viable
> cells?
> We have heard of using liposomes, but wondered if perhaps newer techniques
> have emerged.
>
> Thanks.
>
> Ray Hester
> Univ. of South Alabama
> Mobile, AL 36688
>
> rhester at jaguar1.usouthal.edu
>
>
>
>
>



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