Streptavidin conjugates for MHC-I tetramers - resend with e-mail address in signature

We need to conjugate a home-made MHC-I tetramer to a fluorochrome other
than PE or APC. We routinely biotinylate our MHC-I heavy chain and then
conjugate to streptavidin-PE or streptavidin-APC purchased from
Molecular Probes or BD.  
 
However, we have a population of CTL that can potentially simultaneously
bind 3 tetramers refolded with slightly different forms of the same
epitope peptide. We have already demonstrated simultaneous binding of
the PE and APC-conjugated tetramers. 
 
For this reason, we would like to conjugate the third tetramer to
something like FITC, PerCP, PE-Cy5, PerCP-Cy5.5 or APC-Cy7, or even one
of the Alexa Fluors. We have a FACSCanto cytometer.  Can anyone
recommend from experience one of these streptavidin-conjugated
fluorochromes over the others for this purpose? 
 
Issues that come to mind are spectral overlap/compensation, cost,
stability, non-specific binding, and possibly even steric interference
issues?
 
FITC would be the simplest, but it might be rather dim. The PerCP
conjugate from BD is twice the price of the PE-Cy5. 
 
Thanks in advance for any advice.
 
Simon F. Lacey Ph.D.

Assistant Research Scientist
Laboratory of Vaccine Research
Beckman Research Institute of the City of Hope
Fox South Bld.
City of Hope National Medical Center
Duarte CA 91010-3000

slacey@coh.org <mailto:slacey@coh.org> 
  


"EMF <COH.ORG>" made the following annotations.
------------------------------------------------------------------------------
SECURITY/CONFIDENTIALITY WARNING:  This message and any attachments are intended solely for the individual or entity to which they are addressed. This communication may contain information that is privileged, confidential, or exempt from disclosure under applicable law (e.g., personal health information, research data, financial information). Because this e-mail has been sent without encryption, individuals other than the intended recipient may be able to view the information, forward it to others or tamper with the information without the knowledge or consent of the sender. If you are not the intended recipient, or the employee or person responsible for delivering the message to the intended recipient, any dissemination, distribution or copying of the communication is strictly prohibited. If you received the communication in error, please notify the sender immediately by replying to this message and deleting the message and any accompanying files from your system. If, due to the security risks, you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender.
==============================================================================