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 purification for adherent mammalian cells


  • Please log in to reply
2 replies to this topic

#1 hidayahcellculture

hidayahcellculture

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 39 posts
0
Neutral

Posted 02 September 2013 - 09:35 PM

Hello everybody

hope u all are doing well

 

my Question:

 

To lyse, for adherent cells, after washing with cold PBS, then add RIPA buffer.

Which one come first? do we need to trypsinized the cells first to detach, and after that we add RIPA ?

 

I thought that we need to detach the cells first, is it true?



#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,242 posts
336
Excellent

Posted 03 September 2013 - 02:28 AM

You can lyse the cells directly in the dish if you wish, and then perform some sort of protein quantitation (e.g. Bradford or BCA assays).

 

You can also lift cells with trypsin or by other detaching methods and then lyse either based on cell number per volume or in a defined volume and then perform quantitation as above.



#3 hidayahcellculture

hidayahcellculture

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 39 posts
0
Neutral

Posted 03 September 2013 - 07:31 PM

ok moderator. thank you very much






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

©1999-2013 Protocol Online, All rights reserved.