putative protein crystals?
_
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
xaravich ivan
Sent: Monday, November 01, 2010 9:51 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Crystal gel band
Hi everyone,
I have grown some crystals after micro-seeding starting from
run on the gel
lanes with about the same amount of crystalline volume as your putative
protein crystals?
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
xaravich ivan
Sent: Monday, November 01, 2010 9:51 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Crystal gel band
to get enough S/N for a good experiment?
Good luck!
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
Jim Pflugrath
Sent: Tuesday, November 02, 2010 10:19 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Crystal gel band
It reads like you need to run a lane or two
Hi Ivan,
there are several tests (e.g. Izit dye, crush test) you can do discern
protein from salt crystals but what was always very informative to me
(and certainly in the case of complexes) is a silver-stained SDS-PAGE
gel of the crystals using the following protocol:
- select a drop
Thanks all of you for your answers. As you guessed I was doing coomassie
staining from single crystal (0.10-0.13-0.05).Fortunately I had lots of not
so great looking single crystals from similar drops and I took about 10-12
of them.Then I could see a faint band where I was expecting!
Thanks
Hi everyone,
I have grown some crystals after micro-seeding starting from thin-small
needles from needle-clusters. These crystals are larger in size than the
needles but are comparable to the shape and don't look like salt crystals.
But I cannot see the bands( its a complex) in the SDS-PAGE.I do
Hi ivan,
The detection senstivity of proteins depends on staining technique, You have
not mentioned about staining, whether silver or Coomassie. The silver
staining is more sensitive than coomassie, typically you can see 10-50 ng of
a protein in silver staining. It does vary, however, with the
I have grown some crystals after micro-seeding starting from thin-small
needles from needle-clusters. These crystals are larger in size than the
needles but are comparable to the shape and don't look like salt crystals.
But I cannot see the bands( its a complex) in the SDS-PAGE.I do not have a
