Tricine for elecrophoresis
alkaiser at er5.rutgers.edu
Thu Sep 29 11:33:17 EST 1994
In <36ei9d$fnb at thot.u-strasbg.fr> nussbaum at neurochem.u-strasbg.fr writes:
>In article <fsco-280994095748 at dalessandro.nci.nih.gov> fsco at helix.nih.gov (Francesco D'Alessandro) writes:
>>I am looking for a protocol to make and run a Tricine gel to resolve and
>>isolate a 20KDa protein.
>>Do you have any, or do you have a reference? Does this kind of gel present
>>any problem in performing westerns?
>>Thank you very much.
>I have checked the old handbook of my wife; I think the reference is
>correct; it concerns gel electrophoresis in the presence of tricine
>which worked in my hands ; I did also Western blots with WGA (lectin)
>detection without problems.
>Reference: Schägger et Von Jagow , Analytical Biochemistry , vol 166,
>pages 368-379 (1987)
This reference is correct. You can also check in the current Sigma
catalog p. 1757. Sigma carries a low molecular weight marker kit that
is to be used with this protocol. If you order these markers they send
you the Schagger protocol. However, if you are looking to resolve a
20KD protein I don't think that you need this protocol. Based on what
I know of the tricine gels, they are mainly used for very low MW
proteins and peptides (I use them for a protein with a MW of 3500).
The tricine is there mainly to break up the SDS micelles which form
and can be mistaken for low MW protiens (I guess they stain with
silver stain). If you are looking for a 20KD protein this should not
be a problem. I forget where I read this about the micelles so if
anyone knows any info about this I would appreciate hearing from you.
Hope this helps,
Alan Kaiser alkaiser at eden.rutgers.edu
Graduate Program in Micorbiology and Molecular Genetics
Rutgers University New Brunswick,NJ
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