Inclusion Bodies in E. coli

[CK Wen]

spriley at eclat.uccs.edu (S. Riley) wrote:
>Hello!
>
>Also after isolation of another enzyme (GSA-AT) from a size filtration 
>column in a trycine buffer, pH 7.9, we run it over a DEAE column using 
>the follwing procedure: 0-40% NaCl over 192 mls (the size of our fraction 
>collector). Unfortunately we cannot find any activity after the DEAE and 
>are not sure what has happened to our protein. If you know of any good 
>DEAE procedures please let me know.


Dear Sean:
 
  Before you ran the DEAE-column, did you pre-cycle DEAE ( is it DEAE-
cellulose or sephadex or any other matrix??)  By the way, what was the
size(length and radius, or volumn) of your DEAE-column that you elute
with 192ml of buffer? Also, what was the conc of trycine buffer? Some
buffer may change or reduce the binding capacity of DEAE-column.  If these conditions were not specified, it would be difficult to answer
your questions.

Regards,

Chi-Kuang