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Methanol Protein Precipitation Protocol. The protein precipitate should now pellet to the bottom of the tube. It is suitable for precipitating very dilute proteins and proteins from solutions containing detergents. Efficient protein removal is achieved by mixing the bio-fluid sample with 35times the volume of a water-miscible solvent such as acetonitrile ACN methanol MeOH or a mixture. Protein precipitation is widely used in preparing LCMS samples for bioanalysis.
Four volumes of methanol were added to one volume of the protein sample and the mixture was vortexed. Remove supernatant and add 09-15mL of 100 acetone to wash pellet. To 100 µL protein sample 100µg protein in a 15mL eppendorf tube. Materials Required Cold -20C acetone a volume four times that of the protein samples to be precipitated Centrifuge tube made of acetone-compatible polypropylene and able to hold five times the sample volume g required Protocol 1. Add 110 volume of 3M sodium acetate pH 52 final concentration of 03 M and mix well. 8 Vortex vigorously and centrifuge.
Mix and keep on ice for 45 minutes to 3 hrs cold room 3.
2211 Chloroformmethanol precipitation Experiments were performed at room temperature. MethanolChloroform Protein Precipitation Method can be scaled up or down. To 200 µl protein sample add 480 µl MeOH filtered 160 µl CHCl 3 filtered add under the hood. Centrifuge at 4 o C for 30-45 minutes. Protein precipitation PPT has widely been used for bio-fluid sample preparation for LCMSMS research analysis1. Methanol Precipitation of Proteins.