PRO-PREP™ Protein Extraction Solution
• Simply extract protein from all kinds of cells and tissues
• Monomeric formation is made by ionic detergent
– Effective protein separation is possible for western blot analysis
– Useful for protein molecular weight measurement
• Absorbance hindrance is not contained to eliminate error rate
• Protein stabilization buffer is included for protein stabilizing
• 5 kinds of protease inhibitor included to minimize protein degradation
• Only takes 20~30min for protein extraction
PRO-PREP™ Protein Extraction Solution can be simply extract protein from all kinds of cells and tissues. The kit contains 5 kinds of protease inhibitors so it is possible to extract very highly purified proteins. Usually detergent used in protein extraction consists of both hydrophobic tails and amphiphilic molecule and hydrophilic head. The two parts are joined to form a micelle. It is solubilized protein forming a lipid detergent mixed micelle and transmembrane protein forming a protein lipid detergent complex. The extent of micelle formation is termed as CMC(critical micelle concentration). Which is important for high efficiency as well as high purity of protein extraction. CMC is influenced by pH, temperature, ionic strength, multivalent ions of organic solvents, purity of detergent and so on. Depending on ionic characteristics, a detergent can be categorized as ionic detergent, non-ionic detergent, and Zwitterionic detergent.
Ionic detergent can be further classified into either cationic detergents. These are highly denaturant which have a specific property to isolate protein as a monomeric form and so often used in western blot analysis and measurement of molecular weight. Non ionic detergent such as Triton X-100 are less protein denaturant and often employed in protein interaction. Zwitterionic detergent such as CHAPS have both negative and positive charge head at the same time, more effective in protein-protein interaction than non ionic detergent and its extent of protein denaturation is less than that of ionic detergent. It is very important to select the optimal buffer and detergent for protein extraction.
Fig 1. Add Protease Inhibitor
SDS-PAGE gel Electrophoresis
Total protein was extracted with PRO-PREP ™ Protein Extraction Solution, and then electrophoresed on 15% SDS-PAGE gel and stained with coomassie blue dye.
Lane M, PRO-STAIN I Marker; lane 1, K562; lane 2, SNU-1; lane 3, SNU5; lane4, B16; lane 5, rat Heart; lane 6, cow muscle; lane 7, cow subcutaneous fat
Protein is isolated form variety kind of cell and tissues using PRO-PREP™ Protein Extraction Solution. Total protein volume is measured by PRO-MEASURE ™ Protein Measurement Solution. As a result, we got 2 mg protein form 5×106 cell and 8-9 mg protein is extracted form 10mg tissue.