Hello fellow science researchers :)
I need your help really bad !!
I'm trying to make western blot samples with human blood serum.
But when I mix the serum with 5x sample buffer and boil it,
the sample aggregates and becomes unable to load.
so I used EDTA to lysis? the serum.
it wouldn't harden, but when I loaded the sample and proceeded western blotting
it became all blurry and messy.
do anyone knows how to deal with this problem?
SERUM has high salt content and is alot of protein........ about 60-80mg/ml, hope u have consideed that factor while loading sample. Load max. 100microgram for a blot.
Send more details of yr expt so that we can help you better.
Truth seems so closer now......
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