Minute™ Total Protein Extraction Kit for Adipose Tissues/Cultured Adipocytes (20 Preps) – Invent Biotechnologies Inc.
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Minute™ Total Protein Extraction Kit for Adipose Tissues/Cultured Adipocytes (20 Preps)

Catalog Number: AT-022

  • $288.00


Manual & ProtocolMaterial Safety Data Sheets (MSDS)

Adipose tissue, especially white adipose tissue, 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. Isolation and analysis of both white and brown adipose tissue are challenging due to their high lipid and low protein contents. The water-oil emulsion is also notoriously difficult to separate, but our company has developed technology to deal with this issue. A porous filter with unique surface property and pre-defined pore size/thickness along with a specially formulated detergent-free extraction buffer is employed to effectively separate water-oil emulsion derived from adipose tissue homogenate. The extraction buffer has a lower freezing point than oil in adipose tissues and the aqueous phase can quickly be separated from the oil phase by passing the tissue homogenate through the filter. The total proteins isolated are an unbiased representation of cellular proteins in the tissue. The extracted proteins concentration is very high (2-3 mg/ml) compared to other methods.

 

A. SDS-PAGE (10%) profiles of total protein extracted from different adipose tissues. Lane 1, porcine WAT; Lane 2, chicken BAT; Lane 3, rat WAT.

B. Western blottings of extracted proteins from rat WAT. Proteins were separated in 8-16% gradient 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).

 Kit includes:

Items

Quantity

Buffer A (Extraction Buffer)

15 ml

Buffer B (10 X Denaturing Buffer)

1.5 ml

Buffer C (10 X Non-Denaturing Buffer)

1.5 ml

1.5 ml Microfuge Tubes

20 units

Pestles for 1.5 ml Tubes

2 units

Filter Cartridge with Collection Tubes

20 units

Protein Extraction Powder

2 grams

 

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