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Qproteome Plasma Membrane Protein Kit

For efficient isolation of high-purity plasma membrane protein fractions
  • High-purity functional plasma membrane fractions
  • Highly reproducible using a standardized procedure
  • Convenient and fast isolation in an easy-to-use kit format

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.

Cat No./ID: 37601
Qproteome Plasma Membrane Protein Kit
€418.00
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Buffers and reagents for 6 high-purity plasma membrane protein preparations
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.
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Qproteome Plasma Membrane Protein Kit procedure.
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Efficient isolation of plasma membrane proteins.

Plasma membrane proteins were purified from either HeLa or NIH3T3 cell cultures using the Qproteome Plasma Membrane Protein Kit. Cell lysates [CL] and elution fractions [E] were separated by SDS-PAGE and transferred to a nitrocellulose membrane by western blotting. Proteins regarded as markers for different cell compartments were detected using protein-specific antibodies and an HRP-conjugated secondary antibody with chemiluminescent detection.

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).

Features
Specifications
Applications SDS-PAGE, mass spectrometry
Binding capacity/yield 30–100 µg
Fractions isolated One fraction
Sample size 1 x 10e7 cells
Start material Cell cultures

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Kit Handbooks (1)
For fractionation of plasma membrane proteins from adherent cell culture samples  
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