GuCl in SDS Page

Dr A.H.M. Van Vliet AVV2 at le.ac.uk
Sat Nov 23 10:53:34 EST 1996


bijgh at zeus.bris.ac.uk (Jared Head) wrote:
>Hi,
>
>Can anybody tell me a quick way of prepping samples of protein in 
>guanadinium chloride, so that they will run on SDS PAGE.  The GuCl seems 
>to precepitate when mixed with SDS.  I've only got small samples so dialysis 
>will be difficult.
>
>Cheers,
>
>Jared

Dear Jared,

the GuHCl and SDS indeed precipitate. To avoid that, I have followed
a protocol from the QiaExpress manual using TCA precipitation:

1. Dilute sample(s) to 100 µl
2. Add equal volume of 10% TCA, mix, leave 20 min on ice
3. Spin 15 min in microfuge at maximum speed
4. wash pellet with ice cold ethanol
5. dry pellet, resuspend in SDS-PAGE sample buffer and treat as usual

hope this helps

Arnoud




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