I have plasma samples at -80ºC and I want to perform a 2DE analysis of the proteome. Which could be the best method to thaw them? On ice, at 37ºC. In terms of protein stability.
For plasma proteomics, incubate plasma samples on ice for complete thaw. It will take 30 mins to 1 hr, if you have less time, vortex briefly with incubation. At 37 degree, many enzymes activated, so I preffered on ice.
It depends on the molecules you want to measure. Safe way is to leave the tubes lying on the surface of ice and allow them to thaw slowly. Regularly invert them until ice lump disappears, then stick them straight up into the ice. If you are interested in very stable molecules, you do not care much and you can leave them in a rack to thaw at ambient temp. Highly volatile compounds or unstable molecules prone to oxidation require a much more careful approach and inverting to speed the thawing process is better avoided.
I'll continue thawing on ice, I want to measure the protein content, even identify proteins for biomarker discovery so there will be proteins of very low content so I do not want to lose them. Thank you so much both of you
Albert J Vilella For cfDNA, I thaw on ice if I have time, but if not I do a very short incubation and vortex to thaw, but not warm the plasma. Curious to hear what others are doing though.
Edit: Do you mean what methods to use to extract? I just realized I may have read your question wrong.
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