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:
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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