Qproteome Plasma Membrane Protein Kit
For efficient isolation of high-purity plasma membrane protein fractions
The Qproteome Plasma Membrane Protein Kit provides plasma membrane fractions of very high purity in a fast, reproducible, and standardized procedure that uses a standard bench-top centrifuge.
The Qproteome Plasma Membrane Protein Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Performance
Unique bead-based purification technology in the Qproteome Plasma Membrane Kit delivers the highest purity plasma membrane fractions available from a commercial kit (see figure “Efficient isolation of plasma membrane proteins”).
Principle
Integral and peripheral membrane proteins such as G-protein coupled receptors (GPCRs), receptors for growth factors and cytokines, receptor-associated signaling proteins, and ion-channels represent important potential drug targets for research. Obtaining high-purity membrane proteins fractions, free of cytosolic contamination is a challenge. The Qproteome Plasma Membrane Protein Kit provides plasma membrane fractions of very high purity in a fast, reproducible, and standardized procedure that uses a standard bench-top centrifuge.
Procedure
Cells are incubated in a hypotonic buffer and a mild detergent is added to enable homogenization using a needle and syringe (see "Qproteome Plasma Membrane Protein Kit procedure"). A short centrifugation step removes intact cells, cell debris, nuclei and the major organelles. The resulting supernatant contains cytosolic proteins and microsomes — small vesicles (20–200 nm in diameter) formed from the endoplasmic reticulum, Golgi apparatus, and plasma membranes. Adding a ligand that binds plasma membrane proteins and magnetic beads that in turn bind the ligand enables specific precipitation of plasma membrane vesicles. After washing, plasma membrane vesicles are eluted under native conditions and the ligand remains bound to the beads. Starting material for one fractionation procedure is 1 x 107 cells and typical yields are 30–100 µg protein.
Applications
Unlike other methods, there is no need for biotinylation of surface proteins, meaning that all proteins retain their native conformation and full biological activity — making them highly suited for mass spectrometry analysis, receptor assays, cell signaling studies, and other drug discovery procedures. The virtual absence of contaminants greatly facilitates analysis of low-abundance species (e.g., in mass spectrometry analyses).
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