Conjugation/labeling using carbodiimide-HEL

[carlos h. pedemonte]

>I am currently involved in a research project and I need your help!  Do you
>do cojugations, labelings, etc. to proteins using water soluble
>carbodiimides, especially EDC?  If so, or if you know of procedures for
>doing so, would you please take a few moments and either send me your exact
>procedure or a reference to the one you use?  I am especially interested in
>hearing from those of you who are linking amine functions to Asp or Glu in a
>protein.  Please include in your reponse the following:
>-pH used;
>-time of reaction;
>-concentration of EDC or WSCDI used;
>-concentration of protein/labeling agent; and
>-temp used.
>I am posting this in the hope of getting as wide a variety of procedures as
>possible.  I will send to all who repond a summary of the procedures I get,
>as well as a summary of the results of our experiments (NOTE: I am not
>selling anything, nor will I be!).  Thanks!

I have used EDC to bind nucleophiles to carboxylates of proteins. If you
think that my experience can be useful, you can read the methods in the
following papers:

Pedemonte and Kaplan (1986) J. Biol. Chem. 261, 3632-3639.

Pedemonte and Kaplan (1986) J. Biol. Chem. 261, 16660-16665.

Tell me if you need some more information.

Best, carlos


______________________________________________________________________________
carlos h. pedemonte             Phone:   713-743-1228
University of Houston                    713-743-1211
Dept. of Pharmacology           Fax:     713-743-1229
Houston, TX 77459-5515          e-mail: pedemonte at jetson.uh.edu