Whole brain homogenziation for coIP

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Whole brain homogenziation for coIP

Postby newbie17425 » Feb 08 2018 12:18 am

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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Re: Whole brain homogenziation for coIP

Postby relaxin » Feb 08 2018 5:33 pm

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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Re: Whole brain homogenziation for coIP

Postby newbie17425 » Feb 11 2018 11:35 pm

relaxin wrote: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.


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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Re: Whole brain homogenziation for coIP

Postby relaxin » Feb 13 2018 4:25 pm

You need larger homogenizer.
Retired academic researcher. Mention of a specific product does not imply my endorsement of the product. No conflict of interest or guarantee to work on the advice given. Do as I say, not as I do. Not liable to the loss of your valuable samples.
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