What is the best way to get conditioned medium? - (Mar/14/2007 )

Hellow!

I have to plan a protein array with conditioned medium of adherent cells, but I've never prepared a conditioned medium.

I know I have to replace the complete medium by serum free medium after having let grow the cells, but nothing else. Is there "standard" rules to follow?

Thank you very much guys!

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For obtaining cond medium from 3T3 fibroblasts I ' ve made tight confluent layer of cells then wash cells three times to remove serum obtaining media and incubate in medium+ 0.1% BSA ( for holding cell viabilty during non serum period) 48 hours. Yun should exp your incubation time with your cells. If you will add BSA think about purification from it. But think are you need to purify from BSA ( BSA is a good storage reagent for storing protein) But may be you aims are another?

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For obtaining cond medium from 3T3 fibroblasts I ' ve made tight confluent layer of cells then wash cells three times to remove serum obtaining media and incubate in medium+ 0.1% BSA ( for holding cell viabilty during non serum period) 48 hours. Yun should exp your incubation time with your cells. If you will add BSA think about purification from it. But think are you need to purify from BSA ( BSA is a good storage reagent for storing protein) But may be you aims are another?
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It think I don't need to purify from BSA; I've already used my protein array kit with serum as sample (so full of BSA and other proteins) and it caused no problem.

Thanks for these informations! ;-)

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