Dear Liuqing Chen,
for this purpose I designed an experiment to check using SDS-PAGE if
your HA is bound to your protein or not, this way you don't have to
worry about migration on native-PAGE.
It requires the use of HA that bind covalently to Cys, like HgAc, MMA,
HgCl2, AuCl4, PtCl4, PtCl6, or PiPPt. (Not all Pt bind to Cys).
Using this 7 compounds, I usually find several that bind to my protein,
that can be used for co-crystallization or soaking.
more info in this paper https://www.ncbi.nlm.nih.gov/pubmed/20152903
Feel free to contact me if you need further information
Best of luck
Vincent
On 26/04/2017 10:10, Liuqing Chen wrote:
Hello everyone!
i want phasing my crystal by heavy atom derivazation, before x-ray experiment i
did native page to confirmation which heavy atom may bind to my protein. the
attachment is the native page picture, but the protein is hard to enter in the
gel, my protein pI 5.5, the electrophoresis buffer ph7.5, and run at 60mA for
1h. can anyone give me a suggestion to adust the experiment? or estimate
which heavy atom bind to my protein from the gel picture?
sincerely
Liuqing chen
--
Vincent Chaptal, PhD
Institut de Biologie et Chimie des Protéines
Drug Resistance and Membrane Proteins Laboratory
7 passage du Vercors
69007 LYON
FRANCE
+33 4 37 65 29 01
http://www.ibcp.fr