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[Chlamydomonas] Protein Concentration

Sara Cline via chlamy%40net.bio.net (by cline.228 from osu.edu)
Fri Aug 20 13:49:29 EST 2010


Hello Everyone,

  Does anyone have a preferred method for concentrating crude protein in
Chlamydomonas?

  Here is my situation: I am working on concentrating soluble and membrane
proteins (separately) from Chlamydomonas.  That is, I am freeze-thawing my
cells to create a crude separation of the membrane and the soluble
fractions.  I want to concentrate these two fractions because my protein has
a very low abundance.  Once I have concentrated these fractions, I will
perform an immunoblot.

  I am currently attempting to optimize a protocol using TCA precipitation,
but if anyone has another method that they prefer to use, I would really
appreciate the help.  Or, if you have a TCA precipitation protocol that you
prefer to use, I am having difficulty in the solubilization step.  I believe
pH is the primary problem, but I could be wrong and would appreciate any
tips you could give me.

 Thank you in advance!
- Sara


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