truan at cgmvax.cgm.cnrs-gif.fr wrote:
: group and I found them. Nevertheless I would like to know more about the
: detergents used to solubilize the fatty acids. I have to control exactly
: the concentration of fatty acids so Tween 80 or any equivalent have to be
: discarded I think? I have a protocol which uses tergitol NP40 at 1% but
: when I tried it the agar plates looked very bad. Is it possible to use
: others detergents (like triton or anything elese) at low concentrations.
: The other question is about liquid medium. Is it possible to add the fatty
: acids without detergents?
: Thank you in advance for your help!!
:: Gilles ;->
I don't want to sound snotty, but how can you possibly control exactly the
concentration of fatty acids (FA)? The nature of FA itself makes this a
very difficult (read impossible) task. Studies have been done that show that
FA stick to glass, plastic, or whatever. Therefore, there are naturally going
to be concentration errors due to pipetting (or transfer in general). Then
there are the problems of micelle formation, interaction of the FA with the
container being used, and breakdown (epoxidation) of unsat. FA. All these
affect the relative conc. in media.
We use FA extensively in our research. Normally we must titrate to determine
what FA and how much works best for what we want to see. Sorry I can't give
you a more exact answer. I will give you any and all recipies we use if you
so desire. Additionally, there is an excellent handbook by Calbiochem that
gives many specifics for Detergents. They can be reached at
P.O. Box 12087
San Diego, CA 92112-4180
I by no means work for Calbiochem or are associated with them in any way.
It is just a very handy reference.
Please contact me if I can be of any more assistance.
dubear at molbio.cbs.umn.edu