I guess trypsin is not too bad if you proceed with HPLC separation as
peptides in this range are very well separated by C18 RP-HPLC.
Secondly my opinion is, that this problem has to be solved by mass
spectrometry. This is most efficient cause the results are very precise,
sample load is some picomoles and the only thing you need is a fairly goo=
d
mass spec like many groups are using.
Best regards,
Michael
----------------------------------------
Dr. Michael Schrader
BioVisioN GmbH & Co. KG
Feodor-Lynen-Str. 5
D-30625 Hannover
GERMANY
Phone +49-511-53 88 96-12
Fax +49-511-53 88 96-66
E-mail m.schrader@biovision.de
-----Urspr=FCngliche Nachricht-----
Von: Bao-Shiang Lee <boblee@uic.edu>
An: Recipients of ABRF List <abrf@aecom.yu.edu>
Cc: abrf@aecom.yu.edu <abrf@aecom.yu.edu>
Datum: Freitag, 12. Februar 1999 22:40
Betreff: Re: Thank you!
>Dear Maria,
>
>Trypsin is not a good choice. This produces too many short frgaments and
>you can not use SDS-PAGE at all (you can try limited digestion). You can
>figure out which cysteines form disulfide bridges easily because you kno=
w
>the sequence of your protein. You have many choices to do the digestion =
on
>orther residues. Please post a message one more time with your sequence.
>There sre many experts out there to help you. Hope this help.
>
>Best regards,
>
>Bob Lee
>Director of PRL at UIC
>
>On Fri, 12 Feb 1999, Maria Lorenzi wrote:
>
>> Hi, I'm Maria Lorenzi.
>> Thanks for your advises.
>> I can do the experiment but I cannot digest my protein with CNBr becau=
se
>> methyonines are not present in my protein; I can use trypsin for this
>> purpose and probably it will be good.
>> I know the sequence of my protein and that there are six cysteines on =
it
but
>> I would like to know the number of the disulphide bridges; do you thin=
k
that
>> with your method I can know it?
>> Anyway, thanks a lot
>> Maria
>>
>>
>>
>>
>>
>>
>>
>>
>>
>> "Donde hay Fe hay Amor, Donde hay Amor hay Paz
>> Donde hay Paz esta Dios y donde esta Dios
>> no falta nada"
>>
>>
*************************************************************************=
*
>> Dott.ssa Maria Lorenzi
>> Dottoranda in Biochimica e Biofisica
>> Istituto di Biochimica
>> Facolt=E0 di Medicina e Chirurgia
>> Facolt=E0 di Agraria
>> Universit=E0 degli Studi di Ancona
>> Via Ranieri 60131 Ancona, Italia
>> Tel.:+39 71 220 4677
>> +39 71 220 4681
>> Fax: +39 71 280 2117
>> e-mail: mlorenzi@ascu.unian=
.it
>>
>
>