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.
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2 replies to this topic
#2
bob1
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Thelymitra pulchella
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Posted 08 March 2010 - 03:28 PM
Marker hasn't degraded has it? How much did you load? Have you used it before?
#3
mdfenko
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an elder
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Posted 09 March 2010 - 07:50 AM
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.
you should use at least 7.5% but 10% would be better. a gradient (we like 5-15%) might be even better.
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genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
