Chlamydia antibodies

Richard Haugland richard.haugland at probes.com
Sat Mar 23 22:43:17 EST 2002


Often a good solution to the problem of having only a fluorescein direct conjugate
available is to use R-phycoerythrin anti-fluorescein to simultaneously quench
almost all of the green fluorescein fluorescence, switch the emission to that of
R-PE and significantly amplify the signal. Note that the fluorescein fluorescence
below was collected in the R-PE channel and would have been perhaps 10-fold higher
if collected in the fluorescein channel. We have not made tandem conjugates of
phycobiliproteins with anti-fluorescein but this could presumably shift the
emission further out yet.

Figure 7.63 Color-shifting using a labeled anti-fluorescein/Oregon Green antibody.
Jurkat cells were first stained with a primary mouse anti–human CD3 antibody,
followed by fluorescein goat anti–mouse IgG antibody (F-2761), with the resultant
fluorescence detected in the R-phycoerythrin (red-orange fluorescence) channel of
a flow cytometer (blue curve). The weak signal was then shifted to better suit the
R-phycoerythrin channel by the addition of an R-phycoerythrin conjugate of
anti–fluorescein/Oregon Green antibody (A-21250). The resulting signal intensity
is approximately two orders of magnitude greater (red curve) than the direct
fluorescence from the first staining step (blue curve).

[Image]







Gerhard Nebe-von-Caron wrote:

> I actually use Chlamydia as a nice example to demonstrate the advantages for
> multichannel triggering in the microbiology tutorial. I use the SYVA Microtrak
> kit which it is unfortunately FITC labelled but sufficient to count elementary
> bodies by flow cytometry. If you want to use a green nucleic acid stain you
> might want to add a second step antibody of your choice.
>
> Gerhard Nebe-von-Caron
>
> Research Scientist
> Applied Science & Technology Group
> SEAC - Safety and Environmental Assurance Centre
> Unilever Colworth, Sharnbrook, Bedfordshire,  UK - MK44 1LQ
>
> Tel: +44 (0)1234 264822, Fax: +44 (0)1234 222552
> E- mailto:Gerhard.Nebe-von-Caron at unilever.com
>
> -----Original Message-----
> From:   Chance, Tim [SMTP:jtc3 at CDC.GOV]
> Sent:   Tuesday, March 19, 2002 7:17 PM
> To:     Cytometry Mailing List
> Subject:        RE: Chlamydia antibodies
>
> Does anyone know of a source for flurochrome labeled MoAb to Chlamydia
> bacteria. Preferable PI or PercP tagged.
> Tim Chance
> CDC
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