Hello everyone:
Sorry for the off-topic question. I am trying to separate by SDS-PAGE really
big proteins (500 kDa), and lower percentage (3-8%) acrylamide gels do not do
the trick. Based on literature searches, acrylamide-agarose composite gels seem
the way to go. Is anyone willing to share a
Sorry for the off-topic question. I am trying to separate by SDS-PAGE
really big proteins (500 kDa), and lower percentage (3-8%) acrylamide
gels do not do the trick. Based on literature searches, acrylamide-agarose
composite gels seem the way to go. Is anyone willing to share a protocol?
I
Phil
OK I admit I didn't delve very deeply into the code, but looking at the
printer output it obviously does do a linear fit to the Wilson plot to
get an overall B scale. However, looking at the man page (if all else
fails read the documentation!) I see that it does say that this is only
used
You're right. The smoothed I is used for the Truncate procedure,
though it is difficult in the very low resolution bins. Also it
doesn't allow for pseudo-translations, which it should.
As you say, the linear fit is only used to put data on a very rough
absolute scale. This isn't necessary,
If your protein precipitates at low ionic strength you may want to establish
what concentration of salt does it tolerate (by slowly diluting a fraction
of your pool with water just until you see precipitation) then try to adjust
your cation exchange conditions so that you load with enough salt to
Also, for very large proteins you should not boil the samples - if you must
heat, just go to 50-60C for a few minutes. You also may have better luck
with CTAB-PAGE rather than SDS-PAGE (CTAB tends to smear less, for some
reason).
Artem
-Original Message-
From: CCP4 bulletin board
Phil
Squaring the F (i.e. the best estimate of Ftrue) that Truncate
estimates does not give you F^2 (the best estimate of Itrue):
F = Integral[0:infinity] sqrt(J) p(J|I) dJ
var(F) = Integral[0:infinity] (sqrt(J)-F)^2 p(J|I) dJ
F^2= Integral[0:infinity] J
You can try TOPS.
http://www.tops.leeds.ac.uk/
Best,
Nian
On Fri, Aug 22, 2008 at 8:57 AM, Buz Barstow [EMAIL PROTECTED] wrote:
Dear All,
Does anyone know of a program that is capable of drawing the secondary
structure of a protein molecule?
I'd really like to be able to take a protein
