ReadyPrep™ Protein Extraction Kit (Membrane I), 50 preps #1632088

50 preps, includes 50 ml membrane protein extraction buffer 1 (M1), 50 ml membrane protein extraction buffer 2 (M2), 25 g protein solubilization buffer (PSB), 30 ml PSB diluent, 1 ReadyPrep 2-D Cleanup Kit (1632130), instruction manual; for 50 extractions of 25–50 mg of cells or tissue

Use the ReadyPrep Protein Extraction Kit (Membrane I) for simple, rapid and reproducible preparation of cellular protein fractions highly enriched in membrane and hydrophobic proteins. Protein isolated using this kit can be used for many applications including 2-D gel analysis, SDS-PAGE, and western blotting.

Isolates proteins containing 1–2 transmembrane domains, membrane-associated proteins, and hydrophobic proteins

Protocol can be used with a wide variety of biological samples

Does not require the use of ultracentrifugation or density gradients

Contains a strongly chaotropic extraction buffer to efficiently solubilize proteins

Includes the components of the ReadyPrep 2-D Cleanup Kit to remove constituents of the extraction buffer that can interfere with IEF

The ReadyPrep Protein Extraction Kit (Membrane I) separates membrane proteins by temperature-dependent phase partitioning using Triton X-114 detergent (Bordier 1981, Santoni et al. 2000). Proteins anchored to the membrane or proteins containing one or two transmembrane domains are efficiently partitioned to the detergent-rich phase. Cytoplasmic proteins partition into the aqueous phase. An insoluble pellet contains more complex membrane proteins. Proteins extracted into the two phases are treated using the included ReadyPrep 2-D Cleanup Kit to remove kit components that interfere with IEF. Pellets are then efficiently solubilized using the strongly chaotropic extraction buffer. The solubilized proteins can then be immediately used in 2-D gel electrophoresis or other applications.

Kit Components

For 50 extractions of 25–50 mg of cells or tissue:

Membrane protein extraction buffer 1 (M1), 50 ml

Membrane protein extraction buffer 2 (M2), 50 ml

Protein solubilization buffer (PSB), 25 g

PSB diluent, 30 ml

ReadyPrep 2-D Cleanup Kit (1632130)

Instruction manual