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| PRO-PREP Protein Extraction Solution (C/T) | |||
DescriptionBy using PRO-PREP(TM), proteins can be simply extracted 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 an amphiphilic molecule and hydrophilic head. The two parts are joined to form a micelle, that 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 its 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 : SDS, LiDS and DOC and anionic detergent. Thus 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. Also non-ionic detergent such as Triton X-100 are less protein denaturant and often employed in protein-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 when extracting proteins.CharacteristicsWhen extracting proteins from cells or tissues, one doesn’t necessarily apply appendix treatment.- Able to minimize protein extraction time into 20-30 minutes. - Protein stabilization buffer can make protein stable. - There is no protein degradation due to freezing or thawing since there is no freezing at -20℃ reservation. - Extracted proteins are stable for more than 6 months when kept in –20℃. - There is no absorbable error because there is no absorbable hindrances of PRO-PREP solution when measuring protein concentration. - Very useful for protein separation in Western blot analysis because ionic detergent turns protein into monomers. - Very useful for protein molecular weight analysis because denature protein into a monomers. - Protein degradation is minimized by adding commonly used protease inhibitor and doesn’t necessarily need to prepare protease inhibitor. Protease Inhibitors in PRO-PREP SolutionPMSF(phenylmethylsulfonyl fluoride) - inhibits serine protease and thio protease - added in a working concentration of 1.0mM (174ug/ml) EDTA (ethylendiamine tetraacetic acid) - inhibits metaloprotease - added in a working concentration of 1.0mM Pepstatin A - inhibits acid protease - added in a working concentration of 1uM (0.7ug/ml) Leupeptin - inhibit serine protease - added in a working concentration of 1uM (0.5ug/ml) Aprotinin - inhibit serine and thiol protease - added in a working concentration of 0.1uM (2.0ug/ml) ApplicationsWestern blot assay, Protein-protein interaction, Immunoprecipitation, etcKit ContentsPRO-PREP(TM) Solution 100㎖Storage-20C (stable at least 1 year) Remarks1) Avoid frequent freezing-thawing. PRO-PREP can be frozen below -20℃.2) Use after dissolve the crystals by PMSF (Protein inhibitor). 3) Bubbles can appears during homogenization. Incubate or centrifuge to remove them.  PRO - PREP By using PRO-PREP(TM), proteins can be simply extracted 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 .
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