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
- - - - -

protein extraction and DNP


  • Please log in to reply
No replies to this topic

#1 kingah

kingah

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 22 June 2009 - 03:36 AM

Hi,
I have been having lots of problems with my samples and reaction I need to test recently so I decided to ask for some help from more experienced people :o
In general the aim of experiment is to check the carbonyl groups content in proteins from photoreceptors outer segments. I just isolate outer segments and have them in PBS solution. Then to extract protein I used Folch's method and got an interphase with proteins. Problems start at this stage :( . I need to dissolve my protein to perform reaction with 2,4-dinitrophenylhydrazone (DNP, it is in acidic solution with ethanol) and then get rid of excess of DNP and re-dissolve my proteins in order to measure spectra. I can also perform reaction with DNP with the pellet I get and then centrifuge my sample, followed by dissolution of proteins. I have tried to dissolve protein in 6M guanidine. I have also tried to use trypsin to dissolve it. Nothing worked.
Could anyone suggest some solutions? I am happy to try different methods.
I would be very grateful for your help.
Thanks,
Kinga




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

©1999-2013 Protocol Online, All rights reserved.