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
- - - - -

Protocol for refolding by dilution


  • Please log in to reply
1 reply to this topic

#1 saad

saad

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 49 posts
0
Neutral

Posted 22 March 2011 - 07:24 PM

Dear all

I've been trying to refold protein and by looking to the literature I found the dilution is a good way for refolding.
I diluted my protein in refolding buffer 40 times then I freeze-dry the sample I want to resusped the protein in different buffers and check if is folded/aggregated etc. Is this the right way to do it beacuse I couldn't find a protocol for refolding protein by dilution.
I know we could use Ion exchange chromatography but we haven't a column suitable to deal with 500-600 ml after dilution, we need high flow rate etc.
Could you help me to find protocols to try refolding a protein by dilution.

Thanks

#2 protolder

protolder

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 293 posts
9
Neutral

Posted 22 March 2011 - 10:40 PM

Dear all

I've been trying to refold protein and by looking to the literature I found the dilution is a good way for refolding.
I diluted my protein in refolding buffer 40 times then I freeze-dry the sample I want to resusped the protein in different buffers and check if is folded/aggregated etc. Is this the right way to do it beacuse I couldn't find a protocol for refolding protein by dilution.
I know we could use Ion exchange chromatography but we haven't a column suitable to deal with 500-600 ml after dilution, we need high flow rate etc.
Could you help me to find protocols to try refolding a protein by dilution.

Thanks

Hola, I canŽt give you any protocol wich would be in any book or perhaps in any of the science forums wich all have protocols. You can dilute as you do quickly or dropping diluting buffer that is similar to dialyze, but if you liophylize you will get a high salt concentration. In other time we dialyze against 20mM NH4CO3H at the same pH, wich evaporates in lyophylization. If I found any protocol of refolding by dilution,IŽll send you. Buena suerte




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

©1999-2013 Protocol Online, All rights reserved.