ProteoExtract® Native Membrane Protein Extraction Kit
For the selective extraction of native membrane protein....
Overview: The ProteoExtract® Native Membrane Protein Extraction Kit (M-PEK) is designed for the isolation of native membrane proteins from mammalian cells and tissue. Rather than separating proteins solely by their intrinsic hydrophobicity, M-PEK extracts proteins from mammalian samples based on an actual association of proteins with cellular membranes. Samples from adherent and suspension tissue culture cells (2–5 × 106 cells) or mammalian tissue (25–50 mg) can be separated as the membrane protein fraction in less than two hours without the need for ultracentrifugation or incubation of samples at elevated temperatures. The extremely mild conditions yield a solution of integral membrane and membrane-associated proteins in their native state. These proteins are suitable for enzyme activity assays, non-denaturing gel electrophoresis, Western blot analysis, ELISAs, assays to examine post-translational modifications of membrane proteins, SELDI-profiling of integral and membrane-associated proteins, and NHS ester labeling of membrane proteins for array detection with dyes or biotin.
Features: - Offers three- to five-fold enrichment of native membrane proteins - Yields proteins in their native, functional state - Allows parallel processing of multiple samples in just two steps
Applications:
Selective extraction of membrane proteins from cell and tissue samples Panel A. HEK 293 suspension cells and frozen bovine liver tissue were extracted either with SDS to yield a total lysate or with M-PEK to yield a membrane fraction and remaining “nonmembranous“ proteins. Protein equivalents of extracted fractions were separated on SDS-PAGE and visualized by Coomassie™ blue staining. The membrane protein pattern (lanes 3 and 6) is clearly distinct from the patterns of both total and nonmembraneous fractions (lanes 1, 2, 4, and 5), indicating the selectivity of the M-PEK extraction. Panel B. Immunoblotting of an equivalent gel using membrane-associated and integral membrane protein markers demonstrates the selectivity of the M-PEK extraction procedure.
Mild extraction conditions are compatible with enzyme assays HEK 293 cells were extracted with M-PEK using the two-step procedure to yield a soluble protein and a membrane protein extract. Volume equivalents of the two fractions obtained from two independent experiments were assayed for endogenous alkaline phosphatase activity. The activity profile not only reveals selective separation of this GPI-anchored, membrane-associated enzyme, but also demonstrates that the assayed component is extracted in an active state.
Enrichment for EGF-receptor (EGFR) HEK 293 cells were extracted with buffered 1% Triton® X-100 to generate a total lysate or extracted with M-PEK to yield a membrane fraction. The membrane extract obtained with the M-PEK Kit demonstrated a 4.5-fold enrichment of EGFR.
Add ProteoExtract® Native Membrane Protein Extraction Kit (Cat. No. 444810) to your shopping cart now:
Overview:
Add ProteoExtract® Native Membrane Protein Extraction Kit (Cat. No. 