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De-freezing FBS...


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#1 cshmech

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Posted 08 May 2012 - 09:41 PM

I'm sorry, if its a dumb question. I have purchased around 500mL of FBS as a stock. I will be using that to make my stromal medium. But I'm not sure how to get my FBS. Shall I de-freeze the whole container, take 50mL and freeze 'em again? Would that alter its properties. Previously I was using 50mL frozen FBS, just warm them up once and continue using them.

What does heat-inactivated means?

Thanks

#2 leelee

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Posted 08 May 2012 - 09:47 PM

There is no such thing as a dumb question (well I suppose there is, but this isn't one). You can't learn if you don't ask :)

Yes, thaw the whole bottle, then aliquot the whole bottle out into 50ml volumes and refreeze. A single freeze thaw will not be a problem. (don't just refreeze the 450ml remaining in the original bottle though, coz this will lead to repeated freeze thaws).

Heat-inactivation: see this interesting article/flyer
https://fscimage.fis...nactivation.pdf

#3 cshmech

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Posted 08 May 2012 - 10:00 PM

So, I should take care I don;t freeze-thaw repeatedly? I aliquot all of the FBS into 50mL tubes, and re-freeze them, I can take one, warm it up and use any time. Did I get this right? Actually this makes sense since I was using an old student's stock previously and all were frozen in 50mL centri-tubes. Thanks for the flyer Posted Image

#4 bob1

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Posted 09 May 2012 - 01:01 PM

So, I should take care I don;t freeze-thaw repeatedly? I aliquot all of the FBS into 50mL tubes, and re-freeze them, I can take one, warm it up and use any time. Did I get this right? Actually this makes sense since I was using an old student's stock previously and all were frozen in 50mL centri-tubes. Thanks for the flyer Posted Image

You got it right.




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