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
- - - - -

removing MHC class I from the cell surface


  • Please log in to reply
3 replies to this topic

#1 leelee

leelee

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 652 posts
52
Excellent

Posted 31 October 2012 - 07:48 PM

Hi all

Does anyone know if it is possible to strip MHC class I molecules from the surface of cells (enzymatically, chemically, whatever...)?

I'm having a hard time finding any information on this, as every search I perform comes up with info on stripping the peptides from the mhc I molecules, but not the molecules themselves.

Fingers crossed one of you can help!

Thanks

leelee Posted Image

#2 Astarte Biologics

Astarte Biologics

    member

  • Active Members
  • Pip
  • 20 posts
2
Neutral

Posted 17 November 2012 - 06:24 PM

Low pH buffer (pH 3.0 citrate) can strip the peptides and beta 2 microglobulin off the surface. The class I heavy chain is not stable without the peptide and b2M so it is endocytosed. But new molecules come to the surface all the time. I used brefeldin A to stop transport and was able to see decreased class I on the surface by flow cytometry but they weren't negative, just shifted to the left.

Hope that's helpful.

#3 leelee

leelee

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 652 posts
52
Excellent

Posted 17 November 2012 - 08:05 PM

Very helpful, thank you!
I've been told something similar just recently by a colleague too, however I think they were a bit confused and so not very clear, meaning I had a hard time finding a paper or the like to verify it. Any chance you can point me in the right direction?

Thanks for the info re BfnA and the shifting of the population to the left. I think that will be ok for my purposes, I'm really just trying to explain a certain population of cells I'm getting rather than worrying about complete removal of MHC I from the surface.

#4 Astarte Biologics

Astarte Biologics

    member

  • Active Members
  • Pip
  • 20 posts
2
Neutral

Posted 17 November 2012 - 09:22 PM

Search PubMed for the T2 cell line AND MHC class I and you should find some helpful papers. They are back in the mid-1990's. Let me know if you have any trouble tracking them down and I can give you specific papers.




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

©1999-2013 Protocol Online, All rights reserved.