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problem in IP


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#1 CarrieX

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Posted 01 January 2014 - 01:41 AM

Hello! When I did the IP, I use protein A sepharose beads. I add a negative control which have noting but the beads. When I boil them in the Laemmli sample buffer for 10min, and did SDS-PAGE and western blot, there are smear bands show from ~30kDa to ~60kDa. I really don't understand what are they!? 

Can anybody help?

Thank you!



#2 GNANA

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Posted 01 January 2014 - 12:22 PM

if you boiled the beads alone in Laemmli the smear may be the BSA that was used to pre-block the beads. 


I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....




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