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
- - - - -

SH-SY5Y cells do not grow

SH-SY5Y geneticin

  • Please log in to reply
1 reply to this topic

#1 flyingbullet

flyingbullet

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 14 May 2012 - 02:09 AM

Hey all, I need help

I transfected SH-SY5Y cells with a protein in a pCDNA vector and selected them using geneticin 250µg/ml.
1) the untransfected control plate died 1-2 days later - on the transfected plates only about 80 % died and the rest looked well.
2) the transfected cell do not grow. they are 2 weeks under selection now and seem to be fewer from day to day.
3) the untransfected cells that are not under selection still grow well, so its not a problem of the medium (AMEM, F12, FBS, Pen/Strep, L-Glu)

Where could the problem be???

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,784 posts
405
Excellent

Posted 14 May 2012 - 02:07 PM

What density are the cells at? Selection only works when the cells are actively growing, this means that you typically need them at less than 70% confluency.





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

©1999-2013 Protocol Online, All rights reserved.