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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

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From: Confocal Microscopy List on behalf of simon walker (BI)
Sent: Wed 3/5/2008 10:17 AM
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Subject: Re: Secondary antibody for 405nm laser line ?


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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

 

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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