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I have non problem with Hoechst, I use it for DNA/Nucleus labeling and it
works very well. It's just that I would like to use the 405 channel to label
something else (not the DNA) with an antibody, so I'm looking for a
fluorescent secondary antibody suitable for 405nm exitation.

Christophe

On Wed, Mar 5, 2008 at 5:35 PM, Asson-Batres, Mary <[log in to unmask]>
wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Out of curiosity, what is the problem you are having with Hoechst?  Or are
> you looking for a 405 marker label to improve specificity?
>
> Mary Ann
> Tennessee State University
> Nashville, TN  USA
>
> ________________________________
>
> From: Confocal Microscopy List on behalf of simon walker (BI)
> Sent: Wed 3/5/2008 10:17 AM
> To: [log in to unmask]
> Subject: Re: Secondary antibody for 405nm laser line ?
>
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> We have used Alexa Fluor 405 (Invitrogen) with limited success.  Our
> experience of it is that it's reasonably bright, but bleaches very quickly
> both under epi-illumination and with confocal lsm.  Mounting with an
> antifade seems to help (we've used Vectasheild), but it still bleaches very
> quickly relative to, for example, AF488.  If someone has found a bright and
> stable 405-excitable fluorescent dye I would be very interested to know.
>  Perhaps Qdots maybe a possible alternative?
>
> Simon
>
> Babraham Institute
>
> Cambridge
>
> UK
>
>
>
> ________________________________
>
> From: Confocal Microscopy List [mailto:[log in to unmask]] On
> Behalf Of Christophe Leterrier
> Sent: 05 March 2008 15:46
> To: [log in to unmask]
> Subject: Secondary antibody for 405nm laser line ?
>
>
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear list,
>
> I would like to use all four lines of our new confocal for immunolabeling
> (405,488,532,633). For the moment I use Hoechst/Alexa488/Alexa555/Cy5, but I
> would like to have a fourth antibody labeling using the 405 channel instead
> of Hoechst. What secondary fluorescent antibody would you recommend ? Is
> there an Alexa405 or something similar that would be bright and stable ?
>
> Thanks a lot,
>
> Christophe Leterrier
>
> Postdoc
> INSERM UMR641 Neurobiology of ionic channels
> IFR Jean Roche - Mediterranee University
> Marseille, France
>