Membrane proteins extraction

Membrane Proteins Extraction Membrane proteins (MPs), part of biological membranes, play a crucial role in basic cellular structure and functions, including cell integrity, signal transduction, molecular recognition, material transport, and cell-to-cell communication.

Additionally, MPs are the largest category of drug targets. More than 60% currently available therapeutic molecules target one or more MPs. However, there are relatively few MPs with known crystal structures due to the technical challenges associated with membrane protein extraction, solubilisation, and purification.

MPs are divided into two major classes: integral membrane proteins (IMPs) and peripheral membrane proteins (PMPs). IMPs are permanently attached to the membrane lipid bilayers. While, PMPs are temporarily associated with either the lipid bilayer or IMPs by means of non-covalent interactions. Generally speaking, IMPs can be purified by more stringent techniques than PMPs, whose extraction just needs a high PH buffer. Here we provide a protocol for IMPs extraction focusing on homogenate preparation, soluble proteins removal, membrane proteins extraction and detergent removal.

Reagents: PBS: 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4, pH to 7.4.

Homogenate buffer: 250 mM sucrose, 1 mM EDTA, 10 mM Tris-HCl buffer, pH to 7.2, add protease and phosphatase inhibitors cocktail before use.

Blocking buffer: 0.1% (w/v) bovine serum albumin in 50 mM Tris–HCl, 0.15 M NaCl, pH 7.4.

Washing buffer: 50 mM Tris–HCl, 0.15 M NaCl, pH 7.4.

2% Triton X-100 https://www.creative-diagnostics.com/membrane-proteins-extraction.htm