We are trying to use 100% ethanol as a precipitant for Bovine Serum Albumin. The BSA is in a 0.5% concentration and we are using various ratios of ethanol to attempt to precipitate it, such as 1:2 and 1:3 with increasing amounts of ethanol. Literature reports have demonstrated that ethanol is a good precipitant for BSA in plasma. We are using a normal solution of the BSA in water, and the BSA that we use is fatty acid free. When we run an experiment to attempt to precipitate the BSA from this solution using ethanol, the protein does not precipitate upon exposure to ethanol.
Is there some key difference between plasma bound albumin and our fatty acid free albumin that is rendering ethanol unable to precipitate it?
Your EtOH concentration might be too low. See Figure 4 here
Article Effects of alcohol on the solubility and structure of native...
I suggest adding 9 volumes of EtOH. Also, keep in mind that EtOH is hygroscopic, if the humidity in the air is high 100% will rapidly become 95%,
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