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Forgot to wash the wells

western blot

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#1 Artemis2007

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Posted 05 January 2015 - 12:14 PM

Never did this before, but I was extra distracted today and forgot to wash the wells in my SDS-PAGE bis-tris electrophoresis. What kind of artifacts might I see? 



#2 mdfenko

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Posted 06 January 2015 - 04:39 AM

depends. did you see any differences in the way the sample loaded? if not then there should be no problem.

 

sometimes there are "skins" left between the comb and plate which will interfere with sample loading. these are washed (or scraped) out to allow for consistent loading.

 

also, washing wells will remove, at least temporarily, any buffer components of the gel which may leach into the well and prevent proper loading of the well and may also affect migration.

 

one other thing, washing may also remove debris (bits of gel, dust, etc) from the surface of the well. debris will affect the appearance of the bands and may even cause streaking.


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genius does what it must
i used to do what i got paid to do


#3 Artemis2007

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Posted 07 January 2015 - 12:55 PM

Thanks for your reply. The samples seemed to load ok and looked fine when running. My bands are not showing what I expected, but they're clean and distinct, no streaks. I don't see any staining at the tops of the wells, so I guess the protein migrated ok.







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