finding GFP's mass
klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Fri Apr 13 14:54:03 EST 2001
"Emir Khatipov" <ekhatipo at NOSPAMmidway.uchicago.edu> wrote:
:I think your question was mainly about whether there are other "wetlab"
:methods of determination of molecular weight of a protein. In addition to
:SDS-PAGE you used, there is also
That's what my boss is also saying, but this is very wrong.
Native EF has everything to do with charge and almost
nothing with size.
:gel filtration, ultracentrifugation, mass spectrometry, chemical methods,
:like Kjeldal analysis, and others.
Whta's Kjeldal analysis? Never heard of it. (Should have?)
:"Frederic Langlois" <ye.midget.pirates.deleted.my.account at the.seven.seas>
:wrote in message news:eppz6.12265$up4.324633 at carnaval.risq.qc.ca...
:> Hello.. I'm trying to figure out the different ways to calculate a
:> mass for an assignment. In this particular case I want to know the mass of
:> the green fluorescent protein (GFP).
:> 1) The first method I have used is the SDS-PAGE electrophoresis with the
:> range standard markers. I made a standard curve of the standards Rf value
:> axis) and the molecular weight (X axis). I extrapolated the GFP's mass
:> it's Rf value.
:> 2) Then I searched many databanks and found GFP's amino acid sequence. I
:> used this equation to find GFP's mass:
:> mass = [(n1*molecular
:> where 1=glycin; 2 to 20= all the other amino acids
:> n=the sum of the amino acids of each kind
:> 33g/mol=a water molecule's molecular weight (approximately)
:> I substrated the water molecules that are "lost" when the peptidic bonds
:> I converted the molecular weight in Dalton units...
:> 3) what are the other methods I can use to find the mass of this protein?
:> Thanks in advance,
:> Frederic Langlois
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