Hi Wo, In the past I've added 1 mg/mL BSA to a dilute protein solution, then precipitated it with TCA. However, then you'll be stuck with 1 mg/mL BSA in your resulting pellet.
If you're seriously considering the 24 eppis option, wouldn't dialysis be easier? HTH, Irit _______________________________________ From: [email protected] [[email protected]] On Behalf Of WS [[email protected]] Sent: Monday, April 30, 2012 2:22 PM To: [email protected] Cc: [email protected]; WS Subject: Re: Ammonium sulfate precipitation from solutions with high sugar content Hi Pow, long time no see :) My big problem is that there is almost no protein in the sample -> I won't see any bands. I expect about 1mg from 50 ml. I have started with 50ml of fruit juice concentrate, diluted with 50ml water and then added ammonium sulfate until saturation: No visible turbidity, No (visible) pellet after 2hrs @ 4000g (except some ammonium sulfate crystals. Unfortunately, I do not have access to a high speed centrifuge for such volumes. Spinning 24 eppis with 2ml each @ 15.000g in my small benchtop centrifuge might be an option, however. Could it make sense to add some unrelated protein, eg BSA or OVA (that's what I have in my fridge), as co-precipitant (like one does with glycogen or poly-acrylamide for DNA)?. Wo _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
