CelLytic MEM Protein Extraction Kit

The kit offers a fast and convenient method to isolate hydrophobic and raft microdomain associated proteins from cells. The method is based on phase separation and does not require cell membrane isolation. This kit allows for the preparation of an enriched hydrophobic protein fraction for a number of downstream applications involving protein characterization. The separated proteins can be further analyzed by methods including SDS-PAGE, Western blotting, dot blotting, and immunoprecipitation. The kit has been tested on HeLa, HEK-293, NIH3T3, COS, and CHO cell lines. The kit provides sufficient reagents for 80 tests.

Features & Benefits

• Sufficient for 80 preparations of hydrophobic proteins from 10⁶-10⁷ cells.
• Fast and convenient method to isolate hydrophobic and raft associated proteins.
• Does not require cell membrane isolation.
• Compatible with:
  • Immunoprecipitation
  • Gel electrophoresis - Coomassie Brilliant Blue and Silver Staining
  • Dot blot and Western blot

Kit Components

Lysis and Separation Buffer Wash Buffer for Cellytic MEM Protease Inhibition Cocktail for mammalian cells Sodium Chloride, 4M Solution

Extraction of Membrane Hydrophobic Proteins
MAPK
Connexin-43
Caveolin-1
	1	2	3

Figure 1. Specificity of Hydrophobic Membrane Extraction. HeLa cell membrane hydrophobic proteins were extracted and separated from hydrophilic proteins by phase separation, using the CelLytic MEM Extraction Kit (Product No. CE0050). Lane 1- Total cell extract, Lane 2 -Hydrophilic phase, Lane 3- Hydrophobic phase. Three proteins were used as phase separation markers: MAP Kinase (MAPK), a cytosolic protein, connexin-43, a membrane protein and caveolin-1, a raft-associated protein. The three protein markers are detected in the total cell extract. After phase separation MAPK appears in the hydrophilic phase while both connexin-43 and caveolin-1 are extracted to the hydrophobic phase.

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