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
- - - - -

Need help for dissolving proteins from cells

western protein dissolution SDS

  • Please log in to reply
No replies to this topic

#1 jawsII

jawsII

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 06 January 2012 - 02:57 PM

I am isolating protein from a dendritic cell line using TRIzol reagent. During the last step where the protocol says to dissolve my "protein" in 1% SDS my protein doesn't dissolve--I can see a visible large white solid "protein" in the solution. I've tried to keep my protein solution on a heat block for multiple hours at 60C, but it still won't dissolve!

The TRIzol protocol gives me a few options...



Protein resuspension, continued
Poor solubility of the pellet in SDS can occur, because the solubility
of specific classes of proteins differs with different solvents. If the
protein pellet is insoluble in SDS, the following alternative solvents
(Hummon et. al., 2007) may be required to solubilize the pellet:
 1% SDS and 62.5 mM sarkosyl at pH 8.0–8.8
 9.5 M urea and 2% CHAPS, pH 9.1
 250mM glycerol, 10mM TEA, and 4% CHAPS
 2% diethylamine
 10M Urea

Does anyone have any experience using any of these solvents to dissolve protein for later SDS-page gel use?





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

©1999-2013 Protocol Online, All rights reserved.