Description
General description
A convenient kit for the isolation of native integral membrane and membrane-associated proteins, without the need for ultracentrifugation. Extraction is based on association of proteins with cellular membranes rather than on their hydrophobicity. Resulting samples are suitable for use in functional assays, 2-D gel electrophoresis, and other applications.
Membrane proteins represent only one-third of the proteins encoded by the human genome, but they represent more than two-thirds of the known protein targets for drugs. Therefore, approaches to prepare and characterize membrane proteins are of significant interest for drug discovery. However, due to their hydrophobic nature, membrane proteins are more difficult to analyze than soluble proteins. In addition to the intrinsic difficulty of solubilization, membrane proteins are challenging because of their general low abundance. Thus, the challenges for sample preparation of membrane proteins are effective solubilization and selective enrichment, ideally keeping proteins in a non-denatured state.
Note: 1 kit is sufficient for up to 20 sample extractions.
Biochem/physiol Actions
The ProteoExtract® Native Membrane Protein Extraction Kit is designed for the isolation of native membrane proteins from a broad range of mammalian samples, including:
• Adherent tissue culture cells
• Suspension grown tissue culture cells
• Frozen cell pellets
• Tissues
Preparation Note
• Wash Buffer
Store at 4°C.
• Extraction Buffers I & II
The Extraction Buffers I and II can be stored at 4°C for up to 6 month.
For prolonged storage, freeze the buffers in convenient aliquots at -20°C. Before extraction, buffers must be thawed at room temperature (RT). After thawing, mix components by gently shaking or vortexing. Avoid repeated freezing and thawing!
• Protease Inhibitor Cocktail
The Protease Inhibitor Cocktail is supplied in DMSO and can be stored at 4°C up to 6 months. For prolonged storage, freeze the cocktail in convenient aliquots at -20°C. During the sample preparation procedure it must be kept at RT to prevent freezing of DMSO.
The amount of buffer required for each extraction is dependent upon the amount of starting cell material.
Other Notes
Wash Buffer, Extraction Buffer 1, Extraction Buffer 2, Protease Inhibitor Cocktail, and a user protocol.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
PROTEOEXTRACT is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Multiple Toxicity Values, refer to MSDS (O)
