Becky,
We have never tried 200 µg of Ab but our standard protocol is to
use 300 µg. The key, at least in our hands, is to treat the sample, ie.
all reagents, volumes etc., as if 1 mg of Ab were present. We have labeled
monoclonals 5 times and it has always worked.
George
At 05:03 PM 12/14/00 -0600, Becky Kelly wrote:
>We have tried unsuccessfully to label a small amount of Ab (200 µg) using
>the Alexa
>488 kit. (Since the Ab costs $500/200µg, we didn't go with the
>recommended 1 mg! )
>Does anyone have any suggestions?
>
>From: "George F. Babcock" <babcocgf@email.uc.edu>
>To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
>Date: 12/11/00 2:46PM
>Subject: Re: Alexa dye conjugation kits
>
>
>We have had excellent results with the Alexa 350 dye + instructions and
>488 labeling kit.
>We have not tried the 568. The procedure is extremely easy and we
>obtained consistent
>results from batch to batch. We have used FITC kits in the past and the
>labeling
>procedure is similar in degree of difficulty.
>
>George Babcock
>University of Cincinnati
>School of Medicine
>Cincinnati, OH
>
>At 01:22 PM 12/6/00 -0600, Lynn Dustin wrote:
>
>Hello all,
>A colleague wants your honest opinions of the Alexa dye conjugation
>kits(particularly
>Alexa Fluor _488_ and _568_) from Molecular Probes. Reliability, ease of use,
>alternatives to consider. His application is direct dye conjugation to
>monoclonal
>antibodies for fluorescence microscopy. Thanks in advance for your
>opinions and
>your help!
>
>Lynn Dustin, Ph.D.
>Assistant Professor
>Molecular Microbiology and Immunology
>St. Louis University School of Medicine
>1402 S. Grand Blvd.
>St. Louis, MO 63104
>Phone: 314-577-8415
>Fax: 314-773-3403
>Email: dustinlb@slu.edu
>
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