SDS PAGE of LB medium

Marcelo Foti m.foti at biosidus.com.ar
Mon Feb 18 14:41:26 EST 2002


Dear Michael, I used to precipitate LB medium with TCA, in my experience
less you precipitate better you will see your protein (in the  case of a
good expression).
Try to read the samples with 280/260 nm against a blank made with LB to
discard interferences of oligopeptides present in the media.

hope this help you,

Marcelo D Foti

Argentina


                                                                               
                    Michael Witty                                              
                    <mw132 at mole.bio.ca        To:     methods at hgmp.mrc.ac.uk   
                    m.ac.uk>                  cc:                              
                    Sent by:                  Subject:     SDS PAGE of LB      
                    owner-methods at hgmp        medium                           
                    .mrc.ac.uk                                                 
                                                                               
                                                                               
                    02/18/02 03:27 PM                                          
                                                                               
                                                                               




Dear All,
        I am going to try some SDS-PAGE of the medium my E. coli are
growing in tomorrow - to see if my desired protein is secreted into the
medium.  BUT how much should I load?  I am going to TCA ppt some to avoid
salt, but should I expect a large amount of spurious protein or hardly any
at all?  Plan A is to load ppt from 25microlitres per gel lane.
Regards, Mike.





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