b'Highlights:Optimizedformulations maximize protein yield and preserve protein activity (Figure 1)Efficientonly produces minimal cross-contamination between subcellular fractions (Figure 2)Compatibleextracts can be used directly in most downstream applicationsGentleeliminates the need for mechanical cell disruption for most sample typesA Liver tiLsisvueer t(3is0s umeg ()30 mg) B HeLa cHeellLs a(5 c xe l1ls0 (5 x 10 6 )6 )3,000 3,000 1,400 1,4002,500 2,500 1,200 1,200 Mem-PERM Pelmus- PkEitR Plus kitProtein yield (g) Protein yield (g) Protein yield (g) Protein yield (g)2,000 2,000 1,000 1,000800 800 Mem-PERM keimt (o-PriEgRin akli)t (original)1,500 1,500600 600 EMD MillipEoMrDe PMriolltipeoErex Ptrraoctte koiEtxtract kit1,000 1,000 400 400500 500 200 200 Bio-Rad RBeioa-dRyaPdre Rpe kaidtyPrep kit0 0 0 0MembranMeembranCeytosolicCytosolic Total Total MembranMeembranCeytosolicCytosolic Total TotalFigure 1. Improved protein yield using the Thermo Scientific Mem-PER Plus Membrane Protein Extraction Kit. Membrane proteins were isolated from mouse liver tissue and HeLa cells using four commercial extraction kits. Protein yields for membrane, cytosolic, and total fractions were determined using the Thermo Scientific Pierce BCA Protein Assay Kit.Thermo Scientic Sigma Active Motif Millipore CNCNCNCN HSP90CytosolicMAPK Figure 2. Nuclear and cytosolic fractions are obtained with minimal cross-contamination. HeLa cell proteins were extracted with CalcineurinThermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Reagents or with nuclear extraction kits from other vendors. Samples of the nuclear Thermo Scientic Sigma Active Motif Milliporeand cytosolic fractions were analyzed by western blot using antibodies CNCNCNCNagainst common nuclear and cytoplasmic protein markers and visualized SP1 using Thermo Scientific SuperSignal West Pico Chemiluminescent Substrate. Nuclear fractions produced with the NE-PER kit had minimal to no Nuclear CREB contamination with cytosolic proteins.HDAC1For more information, or to view additional products, go to thermofisher.com/proteinextractionProtein sample preparation thermofisher.com/proteinbiology 15'