Hi RG community,

I want to perform Western Blot on murine adipose tissue. I am currently struggling with some issues to get good protein samples. I usually have been isolating adipose tissue straight from the mice, then minced the tissue with beads or gentleMACS disrupter in protein extraction buffer containing 1% Triton-X. However, I always have contamination of red blood cells which smears my bands and make equal loading impossible. Another issue is fat contamination in the protein sample which may introduce more smearing to the Western Blot.

I am sure some adipose experts out there could help me with some good protocols. Will I need to perfuse the mice to get a good WB sample? Do I need specific buffers?

I look forward to your expertise.

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