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
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5 replies to this topic
#2
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Hmmm, I think it's working
Posted 26 July 2010 - 03:51 PM
I think you want a final concentration of 10 mM.
#3
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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
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Hmmm, I think it's working
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
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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
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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!
