Preparation of cytoplasmic S-100 components

Summary

The contents of a cell contained within the cell membrane. In eukaryotic cells it refers to the portion of the cell membrane outside the inner core that contains structures such as organelles and the cytoskeleton.

Operation method

basic program

Materials and Instruments

Cytoplasm
KCl high salt buffer
Dialysis membrane Centrifuge

Move

1. Measure the volume of cytoplasmic extract and add 0.11 times the volume of 10 x Cytoplasmic Extraction Buffer and mix thoroughly. Centrifuge at 100 000 g for 2 h.2. The supernatant (S-100) is loaded into dialysis tubing and dialyzed until the conductance of the S-100 reaches the same level as the dialysis buffer (KCl 100 mmol/l), and centrifuged at 25,000 g for 20 h. The dialysis tubing is then filled with S-100.
3. Detect the conductance of the supernatant and determine the protein concentration. Dispense into small tubes, snap freeze in liquid nitrogen and store at -80°C.

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