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
- - - - -

Storing conditioned media


  • Please log in to reply
3 replies to this topic

#1 mhum

mhum

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 07 May 2009 - 11:45 AM

Hi,
I'm working on conditioning media from prostate cancer cell lines (DU145 and PC3). After 24 hours of incubation, we want to remove the media and store it (frozen?) since there are many tests we want do (invasion assays, migration assays, VEGF/SDF-1/HIF-1 levels to name a few). So far I have conditioned DU145 cells, have collected and aliquoted the samples (4 x 1ml) and stored them at -20C.

Today when I looked at the samples, many were not frozen, some were frozen, and a few were frozen but there was a clear separation of water and media (large white portion with a small pink layer). In anyone's experience, would it be best to flash freeze the samples before storing at -20C? Should they be stored at -80C?

Thanks,
Mhum

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,242 posts
336
Excellent

Posted 07 May 2009 - 04:39 PM

Many of the secreted compounds are quite unstable, you are best to store at -80, flash freezing would be good too, but shouldn't be too much of a problem.

#3 leelee

leelee

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 652 posts
52
Excellent

Posted 07 May 2009 - 04:53 PM

We used to have a similar problem (things not freezing properly) in one of our -20C freezers. Turned out it was a frost-free freezer. As soon as we moved everything to a non-frost free, it stopped happening.

#4 mhum

mhum

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 08 June 2009 - 12:29 PM

We might have solved the problem - since we saw the same thing happening in another freezer that wasn't frost-free. The samples were being stored in a box, and with the lid on, the cold air wasn't able to circulate, which probably resulted in the uneven freezing. So from now on, we'll freeze without a top on the boxes first :(




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

©1999-2013 Protocol Online, All rights reserved.