Posted 05 November 2004 - 01:26 PM
Hi - if someone can help I will be forever in debt! I work with rhodopsin (GPCR) it's 40kd and seems to be getting stuck up in the gel. I use a 10% gel, and have tried using biorad precast gels too. it does not migrate well but all other proteins i have tried do (BSA, C-MYC etc, cell lysate) Is it aggregating? Am i getting dimers/trimers? It appears to be barely running at all...maybe 10% of the way in the gel (corresponding to 200kd on the ladder) the ladder rund fine but my proteins do NOT! I have made all reagents fresh several times and keep getting the same problem....I am not boiling the protein...(tried that too - but that leads to aggregations of membrane proteins)...nothing seems to work! ANY SUGGESTIONS? PLease post here and email if you feel like it... thanks!!!!