Whole brain homogenziation for coIP

Go to the profile of Chinmaya Sadangi
Feb 08, 2018
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3

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.

3 Comments

Go to the profile of relaxin
relaxin 5 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.

Go to the profile of Chinmaya Sadangi
Chinmaya Sadangi 5 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?

Go to the profile of relaxin
relaxin 5 months ago

You need larger homogenizer.