Thank you very much for your answer! I will try the centrifugation step, this sounds reasonable! What kind of centrifuge do you use? Normal bench top for 1.5-2ml Eppendorf tubes? It's just because of the rotor size.

Dear Kristina, indeed,y to avoid contaminating your protein sample with the fat. What we do after the centrifugation step is carefully removing the 'fat-cake' on top of the sample by a P1000 pipet. Usually that's enough. Another option is to do an additional precipitation of your proteins, although that might give some protein loss.

Dear Dinh-Toi Chu, I extracted protein from mouse BAT tissue for WBs using a buffer from Thermoscientic callled T-PER and following the method that you mentioned above. I can see the protein separation after I did a SDS gel and stain it with coomassie. My problem in the WB is  that when I detect beta actin in my sample, I get two sharp bands instead of one, and none of these two bands correspond to the size of beta actin. I have repeated it and I got the same. Can you think of something that is causing this in my sample? Thanks a lot.

Hi Maricela, 

I detecetd beta-actin in my samples (white adipose tissue) and I did not see two bands.  I have a good detection. Try to run gel with your BAT samples and in one well run other tissue sample and see if you have the two sharps bands...

good luck

Hi Silvia,

How is your protein extraction in the adipose tissue?

Dose the centrifuge process recommended by Dr Dinh-Toi Chu work for you?

Thanks