Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Acetone precipitation and SDS removal

SDS Protein Sampe preparation Acetone precipitation Proteomics

  • Please log in to reply
3 replies to this topic

#1 Antonio Sillo Rosato

Antonio Sillo Rosato

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 25 September 2014 - 09:45 PM

Hi to all,

I'd like to perform an acetone precipitation in order to wash high concentrations of SDS (specifically i have a solution of 10% w/v of SDS) from a protein sample. i'm wandering if it is proper to perform the last centrifugation step (after the incubation of the protein sample in acetone @ -20°C) at room temperature in order to reduce the precipitation of SDS with the protein.

Thank you!



#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,785 posts
132
Excellent

Posted 26 September 2014 - 06:54 AM

(this is a copy of my response at a different forum):

 

i'm not sure if this is a good method to remove protein from a solution of sds. are you also trying to strip sds from the protein?

acetone precipitation of proteins is much more effective at lower temperatures.

you can remove sds by dialysis (and also strip sds from the protein if you include triton x-100 in the dialysis buffer).

you should also be able to remove protein from sds solution (and from the protein) with methanol.


talent does what it can
genius does what it must
i do what i get paid to do

#3 Antonio Sillo Rosato

Antonio Sillo Rosato

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 26 September 2014 - 07:46 AM

thank you. what do you mean with remove sds with methanol?



#4 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,785 posts
132
Excellent

Posted 06 October 2014 - 04:27 AM

thank you. what do you mean with remove sds with methanol?

methanol will precipitate protein. it will also keep sds in solution and strip sds from the protein.


talent does what it can
genius does what it must
i do what i get paid to do





Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.