Suspension cells could be also transfected while in suspensio
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Hy Syed,

Suspension cells could be also transfected while in suspension state and 
immobilized on the coated coverglass immediately before imaging. Yes, antibody 
based FRET works but it is much harder to get impressive and accurate 
quantitative data with the antibody based FRET, as the FRET signal is usually 
weak. However, if you would be happy with the 'YES FRET' and 'NO FRET' readout, 
then the antibody based approach could be OK.

Cheers,

Vitaly
+1-301-515-7833




________________________________
From: "Gelman, Laurent" <[log in to unmask]>
To: [log in to unmask]
Sent: Tue, December 7, 2010 6:51:26 AM
Subject: Re: FRET using fixed cells

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Hi Syed,

We used cells labeled with antibodies coupled to Alexas for FRET with success. 
The Alexa pairs we used were 488&555 and 568&633.

Best regards,

Laurent.

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On 
Behalf Of Syed Khundmiri
Sent: vendredi 3 décembre 2010 16:17
To: [log in to unmask]
Subject: FRET using fixed cells

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Hi all,

I am a novice in the field. I use live cells transfected with tagged proteins 
for 

FRET and TIRF microscopy. My question is can one use fixed cells for FRET 
using AlexaFluor antibodies?? If yes, please guide me to some references and 
how to.

Another question is has someone used TAT-fusion peptides labelled with 
AlexaFluor for TIRF and FRET??

Best Regards,

Syed Khundmiri
University of Louisville