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while making western blot samples with human blood serum, aggregation OCCUR

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yerialice
South Korea

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Topic Started by yerialice
on 6/24/2012 23:16 PM   
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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?
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ARGERINE
India

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Posted By ARGERINE
on 6/25/2012 0:35 AM   
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hi
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.



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