TRITC (FITC) labeling

[Robert Reeves]

I am trying to label plasma fibronectin with TRITC.  The Gold book has
a protocol, but so far all I have been able to label is BSA.  Anyone
have any ideas about what I could be doing wrong?

What I've done:

Resuspend TRITC in DMSO (1mg/ml)
Resuspend pFN in Sodium Carbonate Bffr (0.1M)
Add TRITC to FN  (50ul:1ml)

React 8h at 4 degrees

Isolate over g25 collumn.

Check OD 575/280.  Run on minigel and only see free label.  
Does the TRITC have to be made up fresh each time?  Any insight would
be appreciated!
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Robert Reeves                 rreeves at bu.edu
Boston University
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