Whole brain homogenziation for coIP

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Feb 08, 2018
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Hi,

I want to homogenize a whole brain from a P5 or P50 mouse. Currently, I am using a dounce homogenizer to homogenize the brain in Lysis buffer.

However, I am getting a low yield of proteins from the brain. I sometimes see a large chunk after the lysis. I am unclear if that is causing a low yield of protein and how can I homogenize a whole brain without leaving any large chunk of tissue or membrane.

After protein extraction, I follow it up with coIP.

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Newbie 17425

Scientist, Karolisnka

3 Comments

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relaxin 10 months ago

The most effective homogenization tool is Polytron or similar device.

Brain is quite soft and glass homogenizer should work fine. But it requires chopping the brain into smaller pieces before grinding. Glass tissue grinder with tapered end is more effective than that with rounded end.

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Newbie 17425 10 months ago

Thank you relaxin. I tried again to extract protein but this time I had homogenized 1 brain instead of 2 and it worked fine. Any suggestions for homogenizing two brains at the same time?

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relaxin 10 months ago

You need larger homogenizer.