4 replies to this topic

[celvas]

Hi researchers,

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

Thanks

[phdconsult]

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.

[HirubhaiAmbani]

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.

[mdfenko]

you may have caused your samples to aggregate. you can try running them.

[Gangwolf]

HirubhaiAmbani, on 17 May 2011 - 10:16 AM, said:

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.