Detergent (triton) removal
dk at no.email.thankstospam.net
Thu May 9 23:10:36 EST 2002
"Scott J. Coutts" <scott.coutts at med.monash.edu.au> wrote:
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>I have a couple of questions about removal of triton series detergents
>From the research I've done so far it seems that the most common method
>is the use of BioRad's BioBeads SM2. I've read a few newsgroup posts and
>articles talking about extensive washing in acetic acid, acetone, water
>etc. Some people suggest this acetic acid/acetone/water washing as "per
>the manufacturers instructions"... our biobeads say that they're ready
>to use... does anybody know what the purpose of the acetic acid and
>acetone is, and whether biorad has changed the formulation so that this
>washing is no longer needed?
>Also, what about Amberlite XAD? Has anybody used this for removal of
>TX-100 or TX-114? They're also polystyrene beads, so I assume they will
>achieve the same thing. But they're about 10 times cheaper than the
>biobeads. Are they less effective? Do they have less surface area?
>What's the reason that they're so much cheaper? Is there a protocol
>around or can they just be added and incubated as per biobeads? The
>product sheet doesnt really describe their use, other than that they
>should be 'conditioned' prior to use with water.
>Lastly... any other resins/beads that can be used effectively?
There is also lypophilic Sephadex sold by Pharmacia
("LH-20" or something like that).
One thing that makes all the difference is not clear from you
question: Do you need to remove Tritons from protein-containing
solutions or not? The only truly efficent way to get rid of low
CMC non-polar detergents is to bind protein to a small column
(hydroxylapatite or appropriate ion-exchanger), wash well
and "bump" elute. Pierce claims to have a lypophilic sorbent
that has pores small enough to be inaccesible for proteins.
I tried it once and I lost significantl portion of protein but not
all of the detergent.
If the solution does not contain protein, either prepare it
again :-) or run over BioBeads column (they do not requre
any special super-deper cleaning, just a thorough wash
with the buffer).
More information about the Proteins