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Western and sample boiling


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4 replies to this topic

#1 celvas

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Posted 24 June 2004 - 08:14 AM

Hi researchers,

what difference can I hope in the band pattern if I load my protein sample with and withouth boiling?

Thanks

#2 phdconsult

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Posted 24 June 2004 - 01:51 PM

Try it - 99% of the time you will see no difference at all.

It is not that boiling has no effect; the differences between the two samples are on the picogram level as boiling affects only a few proteins.

#3 HirubhaiAmbani

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Posted 17 May 2011 - 10:16 AM

Hey...I have a quick question.

So, I usually boil by samples in mercaptoethanol and buffer for 10 minutes @ 95 degrees. Today, I forgot about my samples and it boiled for about 40 minutes (did not evaporate). Do you think it will work fine?

Thanks.

#4 mdfenko

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Posted 19 May 2011 - 07:04 AM

you may have caused your samples to aggregate. you can try running them.
talent does what it can
genius does what it must
i do what i get paid to do

#5 Gangwolf

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Posted 19 July 2011 - 06:05 AM

Hey...I have a quick question.

So, I usually boil by samples in mercaptoethanol and buffer for 10 minutes @ 95 degrees. Today, I forgot about my samples and it boiled for about 40 minutes (did not evaporate). Do you think it will work fine?

Thanks.


My lab mate forgot his samples on the hotplate over night and his samples turned out fine. I guess it depends on how sensitive your assay is.




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