Description Adipose tissue especially white adipose tissue (WAT) has been recognized as an important endocrine and inflammation organ in addition to its energy storage function. Isolation and analysis of proteins from adipose tissues are increasingly critical for understanding many physiological/pathological conditions. However isolation and analysis of WAT and brown adipose tissue (BAT) are technically very challenging due to their high lipid and low protein contents. The water-oil emulsion present in biological sample is notoriously difficult to separate. Our company has developed a novel technology to deal with this issue. A porous filter with unique surface property and predefined pore size and thickness coupling with a specially formulated detergent-free extraction buffer is employed to rapidly and effective separate water-oil emulsion derived from adipose tissue homogenate. The extraction buffer has lower freezing point than that of oil in adipose tissues and the aqueous phase can be quickly separated from oil phase by passing the tissue homogenate through the filter. The total proteins isolated are un-biased representation of cellular proteins in the tissue. The extracted proteins concentration is very high (2-3 mg/ml) as compared to other methods.

Application 2D gel analysis, ELISA, SDS-PAGE, immunoblottings, immunoprecipitation, enzyme activity assays, and other applications.

Kit components 1. 15 ml buffer A (extraction buffer) 2. 1.5 ml buffer B (10 X denaturing buffer) 3. 1.5 ml buffer C (10 X non-denaturing buffer) 4. 1.5 ml microfuge tube (20) 5. Pestles for 1.5 ml tube (2) 6. 20 protein extraction filter cartridges with collection tubes 7. Protein Extraction Powder (2 g)

Storage: Store Buffer A at 4oC and Rest of the kit at RT

Additional Materials Required Table-Top Microcentrifuge with a maximum speed of 14,000-16,000 rpm.

Important Product Information The use of protease inhibitors is optional. However if downstream application takes significant amounts of time or the protein extract will be stored for longer period of time, addition of protease inhibitor to buffer A is recommended. For determination of protein concentration BCA assay (Pierce) is recommended. For protein phosphorylation studies, cocktails of phosphatase inhibitors need to be added to buffer A prior to use.