Re: [DIYbio] DIY Protein Separation

wouldn't you use protein SDS gel electrophoresis to separate the
protein? or is that too analytical and you want preparative (i.e. you
need a lot of your pure protein)?

I guess how much protein do you need? I seem to recall Josiah posting
on biocurious that he was cleaning up a F/H-PLC to try and get
working... so I presumed he has some experience with them. I've some
HPLC and GC once in a class, and according to wikipedia ion exchange
chromatography is also considered FPLC sometimes if used for protein,
so when I used the Carolina kit for GFP I guess I was doing FPLC :)

These terms are all a bit overlapping, kinda like the act of walking,
whether you have sandals or boots on. A lot of similarity, but
differences around the edges.

normal phase vs reverse phase (is the buffer (mobile phase) polar or
the resin (solid phase) polar?). You can also have different phases
than solid and liquid. You could say that your lungs filter oxygen and
repel CO2, that would probably be something like gas to liquid to
solid... but that might be stretching it for an /easy/ example :)

On Thu, Jul 31, 2014 at 6:09 AM, Mega [Andreas Stuermer]
<masterstorm123@gmail.com> wrote:
> Hi!
> Again to my question, there seem to be little guys dealing with FPLC here.
>
> https://groups.google.com/forum/#!topic/diybio/24PZTzije98
>
> Is there a way to do this cheaply at my university, without buying an FPLC
> machine?
>
> Thanks god those berberine derivatives are all fluorescent. So detecting
> will not be a problem.
>
> What we want to do is: preparing a cell protein extract. Separate those
> proteins. Add the fractions to berberine, let it incubate,  and do
> photometry see which turns it into the derivative we want.
>
> Best,
> Andreas
>
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-Nathan

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