Hi, you could try without heating. Just vortex your sample a few times after adding the sample buffer and then load the gel.
good luck, Daniela > 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 >
