Protocol for extraction of total protein from organs............ to do westerns .....!

D.K. dk at no.email.thankstospam.net
Mon Jan 26 00:07:00 EST 2004


On Sun, 25 Jan 2004 20:22:44 -0600, "exce-lent" <acb at plouto.net> wrote:

>I will appreciate of I could get a protocol for extracting total protein
>from tissues like liver, brain, kidneys, prostate, ovary etc, for doing
>western blot analysis.

Take a piece of tissue into 2% SDS, homogenize, for good measure 
sonicate to break DNA. If even loading from different tissues is 
required, measure total protein by BCA. After that, add DTT
to 100-200 mM final and boil. Not sure if this counts as a 
protocol but that's what I did in the past for some crude 
samples.

DK




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