Caspases,Paracaspases, and Metacaspases Methods and Protocols

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and caspase-7 to self-process fully as compared to caspase-3.

Additionally, better yields are obtained using a slower expres-

sion protocol (less IPTG) for caspase-2 and caspase-6. Refer to

Table 1 for optimal temperature, IPTG concentration and

Table 1
Caspase expression construct

Caspases Residues

Expression

vector Histidine tag

Expression

time (h)

IPTG (mM)

temp. (°C)

UniProt

reference

Caspase-2 truncated

(no CARD)

159–452 pET-23b(+) C-terminal

LEHHHHHH

18 h P42575

0.02 mM

16 °C

Caspase-3 full-length 1–277 pET-23b(+) C-terminal

LEHHHHHH

5 h P42574

0.2 mM

30 °C

Caspase-3 zymogen 1–277 pET-23b(+) C-terminal

LEHHHHHH

0.3–0.5 h P42574

0.2 mM

30 °C

Caspase-6 full-length 1–293 pET-23b(+) C-terminal

LEHHHHHH

18 h P55212

0.02 mM

30 °C

Caspase-7 full-length 1–303 pET-23b(+)

pET-28b(+)

C-terminal

LEHHHHHH

8 h P97864

0.2 mM

30 °C

Caspase-7 zymogen 1–303 pET-23b(+) C-terminal

LEHHHHHH

0.5 h P97864

0.2 mM

30 °C

Caspase-8 full-length

(-YFP/CFP)

1–479 pET28b(+) C-terminal

HHHHHH

5 h Q14790

0.4 mM

37 °C

Caspase-8 truncated

(no DED)

216–479 pET-15b N-terminal

HHHHHH

6 h Q14790

0.2 mM

30 °C

Caspase-9 full-length 1–416 pET-15b C-terminal

LEHHHHHH

5 h P55211

0.2 mM

30 °C

Caspase-9 truncated

(no CARD)

139–419 pET-15b C-terminal

LEHHHHHH

5 h P55211

0.2 mM

30 °C

Caspase-10 truncated

(no DED)

202–521 pET-15b N-terminal

HHHHHH

5 ha Q92851
0.2 mM
30 °C
aThe expression time is based on full-length caspase-8, not on experimental data

Apoptotic Caspases Assays