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Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form.

Analytical Biochemistry

Aminocaproates, Taurodeoxycholic Acid, Sodium Dodecyl Sulfate, Rosaniline Dyes, Protein Denaturation, Polyethylene Glycols, Octoxynol, enzymology, Mitochondria, Membranes, isolation & purification, chemistry, Membrane Proteins, analogs & derivatives, Glycine, methods, Electrophoresis, Polyacrylamide Gel, Electrophoresis, Gel, Two-Dimensional

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      Abstract

      A discontinuous electrophoretic system for the isolation of membrane proteins from acrylamide gels has been developed using equipment for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Coomassie dyes were introduced to induce a charge shift on the proteins and aminocaproic acid served to improve solubilization of membrane proteins. Solubilized mitochondria or extracts of heart muscle tissue, lymphoblasts, yeast, and bacteria were applied to the gels. From cells containing mitochondria, all the multiprotein complexes of the oxidative phosphorylation system were separated within one gel. The complexes were resolved into the individual polypeptides by second-dimension Tricine-SDS-PAGE or extracted without SDS for functional studies. The recovery of all respiratory chain complexes was almost quantitative. The percentage recovery of functional activity depended on the respective protein complex studied and was zero for some complexes, but almost quantitative for others. The system is especially useful for small scale purposes, e.g., separation of radioactively labeled membrane proteins, N-terminal protein sequencing, preparation of proteins for immunization, and diagnostic studies of inborn neuromuscular diseases.

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      Most cited references 16

      • Record: found
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      A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding

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        Cleavage of structural proteins during the assembly of the head of bacteriophage T4.

         U K Laemmli (1970)
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          Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa

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            Journal
            1812789

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