Mouse UCP-I Quantification in Brown Adipose Tissue
Instrument: Precellys® 24 Dual
Sample: Brown adipose tissue
Protocol:
- Precellys kits: CK28 (big ceramic beads)
- Sample : brown adipose tissue (between 30-50mg)
- Buffer : 25mM TrisHCl - 1mM EDTA - 1% Triton X-100 0.5% Na-deoxycholate : 500μl
- Run the Precellys® 24 Parameter at 5000rpm, 2x50 sec., 5 sec. Break
- Western blot against mouse UCP-1 1st antibody: rabbit anti-UCP-1 2nd antibody: infrared (IR)- labeled antirabbit
- Detection of bands with odyssey Infrared Imaging System
Results:
The lanes correspond to 8 different concentrations of total proteins in order to control the resolution by using the infrared imaging system. UCP-1 quantification studies enable us to get more information about body temperature regulation. Results between control and cold-exposed mice remain confidential.
Detection of lanes with Odyssey Infrared System by using IR-labeled 2nd antibodies. The lanes correspond to 10- 30-50-70-20-40-60-80mg concentrations of total proteins.
