Hi,
I use 4X LDS buffer from Invitrogen, I wonder if I can just boil the sample in 95C instead of 70C? will it hurt? Thanks!!
TFC
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#2
fishdoc
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Posted 31 December 2009 - 07:22 AM
tfcheng, on Dec 30 2009, 01:04 PM, said:
Hi,
I use 4X LDS buffer from Invitrogen, I wonder if I can just boil the sample in 95C instead of 70C? will it hurt? Thanks!!
TFC
I use 4X LDS buffer from Invitrogen, I wonder if I can just boil the sample in 95C instead of 70C? will it hurt? Thanks!!
TFC
I've boiled for 10 min every time I've used the stuff, and it works fine. I haven't compared it to 70C to know if it's better, worse, or equal.
#3
mdfenko
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Posted 31 December 2009 - 07:43 AM
the reason to use 70C instead of 95C is to prevent aggregation of the protein. this can happen if you incubate for too long at 95C or higher (it is worse for some proteins than others).
65-70C is a safer incubation temperature, especially if you have to incubate for longer periods.
this goes for sds as well as lds.
65-70C is a safer incubation temperature, especially if you have to incubate for longer periods.
this goes for sds as well as lds.
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