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Protein extraction from mouse prostate


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#1 shyn

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Posted 04 November 2010 - 12:31 AM

Hi all,


Does anyone has experience isolating protein/ RNA from mouse prostate? As the organ is really small, I'm concerned about how I should go about extracting the protein (for Western) and RNA (for PCR) with minimal loss.

Also, it would be great if anyone out there is able to share useful tips on how to isolate the mouse prostate.


Thanks!

#2 gfischer

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Posted 04 November 2010 - 07:15 AM

I'd recommend trying simple homogenization in RIPA lysis buffer with protease and phosphatase inhibitors. I've used that for rat DRGs (about 2mg) and usually get 80-100ug of protein per sample.
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#3 shyn

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Posted 07 November 2010 - 05:44 PM

I'd recommend trying simple homogenization in RIPA lysis buffer with protease and phosphatase inhibitors. I've used that for rat DRGs (about 2mg) and usually get 80-100ug of protein per sample.


Hey gfischer,

Thanks for the reply. What type of homogenizer are you using? The electrical type, or a dounce homogenizer? I'm worried about tissue loss mainly.

Edited by shyn, 07 November 2010 - 05:45 PM.


#4 gfischer

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Posted 08 November 2010 - 01:29 PM

I've used dounce and motorized homogenizers, and both of them work fine. Try not to use too large a volume. 100-200uL is a good start (you can always dilute the sample if your concentration is too high, but if it's too low, you're stuck doing precipitation). Depending on what size probe you have, it may be difficult to use a motorized homogenizer into a small enough tube.
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