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Western blot narrow bands


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#1 kedong

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Posted 07 June 2012 - 04:10 PM

Hi everyone,

I do a lot of western blots in my lab and have thus far been problem free. But in the past week or 2 I've been getting some narrow bands or an hour glass figure for my lane when I run my gels. The standard protocol for my lab is to use 100V to run the samples through the stacking gel then once the ladder begins to separate change the voltage to 200V and run it until the dye is ~1cm from the bottom the gel. After that do either a overnight transfer at 20V or use the Bio-rad fast transfer machine for much faster transfer (40min).

Here's a photo of a 15% gel I ran:

Posted Image

Thanks,

Chris

#2 mdfenko

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Posted 15 June 2012 - 08:30 AM

i can't view your image

are you running the same samples as before you saw the change?

you may have higher salt or lipid or detergent or nucleic acid concentrations in some samples.
talent does what it can
genius does what it must
i do what i get paid to do




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