Besides the K-SDS precipitation it can also be the problem of membrane
proteins generally having a tendency to aggregate when boiled with
SDS. Try to just add the Laemmli buffer and load the gel without
boiling. This is the general procedure for membrane proteins.
Poul
On 18/12/2008, at 06.20, Artem Evdokimov wrote:
Is your phosphate a Na-K phosphate? If it is, then you're probably
getting a
K salt of SDS which is considerably less soluble than Na or Li salts.
Artem
-----Original Message-----
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
Subscribe Ccp4Bb Kn L
Sent: Wednesday, December 17, 2008 9:26 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] LDAO & SDS-PAGE
Hello everyone...
I have a protein solubilised in 1% LDAO, 50 mM Phosphate pH 7.5, 150
mM
NaCl. When I mix it with laemmli buffer and heat at 95C, 3 mins for
SDS-PAGE, it aggregates (as in when you mix GuHCl with SDS) and
can't be
loaded onto the gel.
Does anyone know why it happens? Thanks a lot!!!
Kien
