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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: Cold Spring Harb Protoc. 2013 Sep 1;2013(9):880–884. doi: 10.1101/pdb.prot073056

TABLE 1.

Optimal conditions for isolating nuclei from a variety of cell types

Cell type [PMSF] in
NIS (µm)a 		Cell density in
NIS (×106/mL)		 Homogenizerb Number of
strokesc 		Reference
Sf9 100 1.6 Dounce 4–6 Ionescu et al. 2006
HEK293-mRyR2d 50 1.5–2 Dounce 4–6
S2 50 2–3 Duall-1 7–9
DT40 50 2–3 Duall-1 7–9 Li et al. 2007
N2a 200 1.5–2 Duall-2 14–16 Kopil et al. 2011
PC12 200 1.5–2 Duall-2 12–14
SH-SY5Y 200 1.5–2 Duall-2 9–12
HEK293 200 1.5–2 Duall-2 14–16
Human B lymphoblasts 200 2–3 Duall-2 10–12 Cheung et al. 2010
Mouse primary cortical neurons 200 2–3 Duall-2 8–10 Cheung et al. 2010
Mouse embryonic fibroblasts 200 1.5–2 Duall-2 12–15 Cheung et al. 2010
a

These values are given only as a reference. The optimum [PMSF] varies depending on the culture conditions and cell type.

b

Duall-1 and Duall-2 refer to Duall homogenizers of weaker and stronger homogenizing strength, respectively. See Discussion for further explanation.

c

The number of strokes to use is very strongly dependent on the particular homogenizer used, as well as on the age of the cell culture.

d

HEK293 cells expressing recombinant RyR (Jiang et al. 2007).