Does anyone have a preferred method for concentrating crude protein in
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!