Re: Antibody Conjugation Vendor and Alexa633

From: Howard T Petrie (h-petrie@ski.mskcc.org)
Date: Fri Feb 01 2002 - 16:41:02 EST


Having spent many years conjugating many antibodies to
phycobiliproteins, I can understand why you wouldn't want to do it
yourself (although the results are ALWAYS better).  For the last
couple of years, however, we've been using Alexa633 instead, and I
have to say that even using the straight-out-of-the-box protocol
supplied by Molecular Probes, the results are quite good (if you have
enough antibody to spare and want to test a couple of chrome:Ab
ratios the results can be even better).  I highly recommend giving it
a try, it takes about one hour of hands-on time and about three hours
in total.

I should mention that Alexa660 works beautifully as well for flow, in
fact better than 633 (I'd love to hear somebody at either MP or one
of the equipment manufacturers explain why this should be the case;
neither the optics nor the spectra would predict it).  In fact after
years as a dye laser loyalist, we now use HeNe exclusively, since we
can simultaneously excite and detect A633 and A660 instead of APC and
TR.  Given the utter simplicity of conjugation of the Alexa dyes,
it's worth considering.

>	To All:
>
>	I am in need of getting two monoclonal antibodies conjugated to APC.
>	If you have had this done by some company/lab and liked the result
>would you please let me know.
>
>	thanks
>
>	Jim Houston

--


Howard T. Petrie, Ph.D.
Head, Laboratory of Developmental Immunology
Memorial Sloan-Kettering Cancer Center
Box 341, 1275 York Avenue
New York, NY 10021
phone (212)639-2149
fax (212)794-4019



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