pre stained marker issues - (Mar/08/2010 )
Hello,
I am running a 5% gel and am seeing some issues- I transfected HEK293 cells and fibroblasts with my plasmid of interest. I obtained the cell lysates by incubating the cells with RIPA buffer, centrifugation and collecting supernatant.
When I run the gel, the pre stained marker is not separating at all. My protein size is 110KD. I only see the blue line from the samples on the gel but no clean separation of the marker.
Can anybody help me on this issue?
Thank you.
-xyz74-
Marker hasn't degraded has it? How much did you load? Have you used it before?
-bob1-
a 5% gel is neither very selective for a 110kDa protein nor for your markers.
you should use at least 7.5% but 10% would be better. a gradient (we like 5-15%) might be even better.
-mdfenko-