IUBio

Amino acid analysis problems

Achim Recktenwald achimr at home.com
Thu Mar 4 23:20:26 EST 1999


Since it's a membrane protein, it probably has some very hydrophobic patches
that might bind strongly to lipids or similar interfering compounds. Have
you tried delipidation of the protein, e.g., by acetone precipitation,
before the hydrolysis.

Cheers,

Achim


ummanley at cc.UManitoba.CA wrote in message <7bmcau$car$1 at mserv2.dl.ac.uk>...
>
>Sorry.....
>
>I express in the E.coli strain BL21(DE3)pLysS.  Actually I use a 4M urea/
>SDS mixture for solubilization (separately the urea and SDS are
>inefficient at solubilizing my protein, the best detergent is
>dodecylmaltoside.....but its very expensive). The protein is a natural
>E.coli membrane protein, and there is no evidence of inclusion bodies
>(even EM doesn't show anything).
>
>Do you suggest removing the SDS instead of the urea for AAA?
>
>Thanks,
>
>Darren
>
> ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
>{                                                                }
>{ Darren Manley                         Department of Chemistry  }
>{ phone (204) 474-6697                  University of Manitoba   }
>{ fax (204) 474-7608                    Winnipeg, MB., R3T-2N2   }
>{ email ummanley at cc.umanitoba.ca        Canada                   }
>{                                                                }
>{                                                                }
>{        Quantum Mechanics: The dreams stuff is made of.         }
>{                                                                }
> ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
>





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