Bio-Rad ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) - Manual (Page 7)

Section 4
Instructions for Use
Two protocols have been provided. They differ based on
the type of starting sample material – one for cultured cells,
one for tissue samples. Use the protocol that best fits the
experimental sample. Please read the entire protocol before
using the kit.
4.1 Protocol A
4.1.1 Extraction of Cytoplasmic/Nuclear Proteins From
Cultured Cells
Note: Chill CPEB on ice for at least 15 min before
beginning.
1. Transfer cell culture suspension or adherent cells
(1–5 ml) into a 1.5 ml microcentrifuge tube and
centrifuge to obtain approximately 0.05 ml of packed
cells. Remove and discard the supernatant.
2. If the cells have not previously been washed, add
1 ml of ice-cold 1X phosphate-buffered saline solution
to the cell pellet. Pipet up and down several times or
vortex briefly to resuspend the cells, and pellet the
cells again by centrifugation. Remove and discard the
wash solution.
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