Membrane Protein Extraction Kit
A kit for fast and easy extraction of membrane proteins from eucaryotic cells and tissues
employs a detergent-based temperature-dependent phase separation to enrich membrane proteins from mammalian tissue or cell homogenates. By this simple and rapid procedure you obtain a rather pure, non-denatured membrane protein fraction from any chosen protein homogenate. Most membrane proteins partition to the detergent phase with 80-90% efficiency, while the soluble proteins generally remain in the aqueous phase.
Depending on the starting material, the membrane proteins in the detergent phase will constitute 15-25% of input protein, thus being separated from many contaminating soluble proteins and other water-soluble compounds. Proteins with a very low expression level like Cellubrevin may become accessible to analysis only after enrichment. Abundant proteins like Tubulin or Synaptotagmin 1 can be found in both phases and may need a second round of extraction for complete purification. However, each protein shows individual behaviour depending on hydrophobicity and interactions with other (hydrophobic) proteins.
The membrane proteins isolated by SySy MemEx Kit are suitable for downstream applications like activity assays, immunoprecipitation, proteomic analysis or SDS-PAGE/western blotting. If the detergent should interfere with analysis, it can be removed by protein precipitation (denaturing) or dialysis against 0.5 % CHAPS in a suitable buffer (native; CHAPS can later be exchanged with other detergents).