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. 2020 Jun 5;10(11):e3631.
doi: 10.21769/BioProtoc.3631.

Mouse Adipose Tissue Protein Extraction

Yu A An  1 , Philipp E Scherer  1
Affiliations

Mouse Adipose Tissue Protein Extraction

Yu A An et al. Bio Protoc. .

Abstract

As obesity becomes a global epidemic, the metabolism research field is increasingly focusing on studying the physiological and pathological roles of adipose tissues (AT). However, extracting proteins from AT is challenging due to abundant fat content of intracellular lipid droplets. Several commercial kits for extraction of AT proteins are available, as are protocols (such as the RELi protocol as well as other protein precipitation protocols). The protocols have been introduced to improve the quality and yield of extractions, but these methods either increase the cost or involve multiple steps. Herein, we describe a detailed protocol for mouse AT protein extractions based on our daily laboratory practice. This protocol requires only very common reagents and instruments, and can be completed in 90-120 min and provides good recovery of total protein content. Thus, this protocol is an economically attractive, time-saving and efficient way to extract proteins from the AT.

Keywords: Adipose Tissue; Lipid Contamination; Metabolism; Mouse; Obesity; Protein; Time and Cost Effective.

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Conflict of interest statement

Competing interestsThe authors declare no competing interests.

Figures

Figure 1.
Figure 1.. Mouse fat pads harvested in different locations.
A. Subcutaneous WAT (sWAT). B. Interscapular brown AT (BAT). C. Epidydimal WAT (eWAT).
Figure 2.
Figure 2.. Fat cakes in different samples.
After homogenization and centrifugation, fat cakes, indicated by arrows, are the white lipid layers on top of the aqueous layers in sWAT, BAT and eWAT samples.
Figure 3.
Figure 3.. Representative Western blot images of protein samples extracted from AT.
A total of 15 μg protein in each sample was loaded on an SDS-PAGE gel. Adiponectin and an internal control β-actin in sWAT (left), eWAT (middle) and BAT (right) were probed.
See this image and copyright information in PMC

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