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!
- - - - -

anyone has experience in adhesion assay?

icam lfa adhesion t cell

  • Please log in to reply
No replies to this topic

#1 gyma



  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 146 posts

Posted 20 May 2013 - 11:19 PM

hello everyone I had a problem in adhesion assay.

I was testing the effect of a drug on T-cell adhesion. I coated recombinant ICAM1 in 96-well plate overnight at 4C and blocked it with 0.5%BSA for 1 h at 37C the next day. Then I seeded 5X10^4/well of T-cells (which have been confirmed to express LFA1, the ligand) and incubated the plate at 37C for 2 hrs. Here comes the problem. Most references just say that non-adhered cells are washed away. But how? I added PBS and then removed it immediately. To make sure that remaining cells were really adhering to ICAM1, I washed 3 times. And finally only very few cells remained. How should I wash the wells? Anyone has experience? Please tell me your advice, thank you.

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

©1999-2013 Protocol Online, All rights reserved.