Minute™ Adipose Tissue Fractionation Kit
In average, total cellular proteins account for less than 2% of adipose tissues. Fractionation of adipose tissue
is technically very challenging due to its high lipid and low protein contents. Adipose tissue especially white
adipose tissue (WAT) has been recognized as an important endocrine and inflammation organ in addition to
its energy storage function. Fractionation and analysis of proteins from adipose tissues are critical for
understanding many physiological/pathological conditions. Thewater-oil emulsion present in biological
sample is notoriously difficult to separate. We have developed a novel technology to deal with this issue. A
porous filter with unique surface property and pre-defined pore size coupling with a specially formulated
detergent-free fractionation buffer is the key to fractionate adipose tissue into two fractions: water-soluble
protein fraction containing mainly cytosolic proteins and water insoluble fraction containing mainly plasma
membrane, and organelles such as mitochondria.The buffers used in this kit are free of primary amine,
detergent and reducing agents. Isolated proteins are compatible with all downstream applications including
TMT labeling, enzyme digestion, MS analysis and other applications.
A. SDS-PAGE (10%) profiles of fractionated adipose tissues. Lane 1, water soluble fraction of porcine WAT;
Lane 2, water-insoluble fraction porcine WAT; Lane 3, water soluble fraction of rat WAT; Lane 4, water-
insoluble fraction of rat WAT.
B. Western blottings of fractionated proteins from rat WAT; Lane 3. water solublr fraction; Lane 4 water in-
soluble fraction. Proteins were separated in 10% SDS-PAGE and probed with following cellular protein marker
antibodies: Anti-Na/K ATPase alpha1, a plasma membrane marker(Upstate, clone 464.6), anti-lamin B1, a
nuclear envelope marker (ab16048, abcam Cambridge, MA), anti-ubiquinol-cytochrome C reductase core
protein (abcam, ab 96333) and GAPDH, a cytosolic marker (Sigma). The specific protein bands were
visualized by a substrate Opti-4CN (Bio-RAD).