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cell culture sample preparation for HPLC


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#1 janap

janap

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Posted 09 October 2013 - 11:14 PM

Hi everybody, 

I would like to ask for protocol for cell culture samples preparation for HPLC. I am going to measure antioxidants levels in adherent cell cultures. I need to precipitate proteins and I should do it by 90% acetonitrile + 0.1% TFA (according to my boss).

I just don't not how much acetonitrile to add to cell pellet, how long to vortex the mixture, keep on ice or not, what speed to centrifuge etc.

Second question, can I freeze these acetonitrile samples in -80°C and for how long?

Thank you for any comments!

 

Jana






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