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
- - - - -

Adding L-glutamine to DMEM media w/o L-glutamine


  • Please log in to reply
5 replies to this topic

#1 coco

coco

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 26 July 2010 - 10:22 AM

Hi all!
I'm trying to do an MTT assay using no phenol-red, no L-glutamine DMEM media and will be transferring mesenchymal stem cells from (+)phenol red (+) L glutamine media into this no L-glutamine media, no phenol-red media [b/c phenol red negatively affects MTT results].

I was wondering, how much L-glutamine should I add into the no-L glutamine DMEM? There is 500 ml of media.

Thanks!

-coco

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,584 posts
384
Excellent

Posted 26 July 2010 - 03:51 PM

I think you want a final concentration of 10 mM.

#3 Piersgb

Piersgb

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 41 posts
0
Neutral

Posted 27 July 2010 - 02:22 AM

I'm working with mouse bone marrow cells and the basal media that I use (RPMI 1640 - Invitrogen) that already has L-Glutamine in is at a final concentration of 2mM. Just my two cents!

#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,584 posts
384
Excellent

Posted 27 July 2010 - 04:05 PM

Actually, yes, 2 mM is more what I recall. The bottles of medium with L-glut in them should have the concentration on the label or product sheet.

#5 shldbcrzy

shldbcrzy

    member

  • Active Members
  • Pip
  • 26 posts
0
Neutral

Posted 27 July 2010 - 04:34 PM

does adding extra 2mM of glutamine to the media have any adverse effect ( if the protocol requires only 2mM )?

#6 Piersgb

Piersgb

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 41 posts
0
Neutral

Posted 28 July 2010 - 04:10 AM

It could do, I'm not sure. I guess what the manufactorer provides in the media is a kind of standard/basal amount of L-Glutamine. The best way to figure this out for your cells would be to try adding extra and not adding extra L-Glutamine to your media and having a go at culturing your cells. It's a small factor but the smallest of things can sometimes have the biggest impacts!




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

©1999-2013 Protocol Online, All rights reserved.