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brain and colon tissue homogenate for WB


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#1 topsi

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Posted 16 February 2012 - 12:50 AM

Hello,
I need to do ELISA on mice colon and brain samoles and my protocol asks for homogenization in PBS. My question is, do you think those homogenates will be appropriate for TGF beta WB??
Also, for WB, what would you recommend: tissue homogenization by ultrasonic cell disrupter (on ice) or mortar and pestle grinding in liquid nitrogen?
Thank you!

#2 Inmost sun

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Posted 16 February 2012 - 06:52 AM

you should at least separate cytosolic from microsomal fraction

grinding in liquid nitrogene is appropriate for colonic tissue as exocrine proteases may cause special problems to cellular proteins

#3 topsi

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Posted 17 February 2012 - 12:33 AM

Thanks, I will use the grinding methotd then.
As for the first reply, what exacrliy do you mean by that..?

#4 Inmost sun

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Posted 17 February 2012 - 06:38 AM

differential centrifugation




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