Ammonium sulfate precipitation from solutions with high sugar
(by irappley from scripps.edu)
Mon Apr 30 23:52:18 EST 2012
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?
From: methods-bounces from oat.bio.indiana.edu [methods-bounces from oat.bio.indiana.edu] On Behalf Of WS [novalidaddress from nurfuerspam.de]
Sent: Monday, April 30, 2012 2:22 PM
To: bionet.molbio.methds-reagnts from googlegroups.com
Cc: methods from magpie.bio.indiana.edu; 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)?.
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