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
- - - - -

problems with cell stimulation


  • Please log in to reply
2 replies to this topic

#1 kfrenzel

kfrenzel

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 20 May 2004 - 01:29 PM

When I stimulate cells with agonist and look at the resulting tyrosine phosphorylation, the signal in the 0 timepoint (unstimulated) is always equal to the signal in the "stimulated" timepoints. I have tried serum starvation for 24h or 48 or even 72 hrs without reducing the signal at 0 time. Any other suggestions?

#2 chaos

chaos

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 23 May 2004 - 06:16 PM

The stimulation time is also important. I used EGF 100ng/ml to stimulate HCT-116 for 10 min, then i got a thick band in WB, but got no result for 15 min just as unstimulated.

#3 postdoc2130

postdoc2130

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 35 posts
0
Neutral

Posted 25 May 2004 - 04:27 PM

Add tyrosine phosphatase inhibitor and do a time course and dose-response studies.




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

©1999-2013 Protocol Online, All rights reserved.