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Caspase 3/7 Activity

LH Li He
TS Tejasav S. Sehrawat
VV Vikas K. Verma
AN Amaia Navarro-Corcuera
GS Guneet Sidhu
AM Amy Mauer
XL Xin Luo
TK Tomohiro Katsumi
JC Jingbiao Chen
SS Soni Shah
JA Juan Pablo Arab
SC Sheng Cao
HK Hamid Kashkar
GG Gregory J. Gores
HM Harmeet Malhi
VS Vijay H. Shah
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The assay was performed as described by us previously (Malhi et al., 2006). Cells were plated in 96-well plates (Corning Inc., Corning, NY, United States). Caspase activity assay was performed using the commercially available Apo-ONE homogeneous caspase 3/7 assay (Promega Corp.) according to the manufacturer’s instructions. Briefly, this assay involves cleavage of a profluorescent caspase 3/7 consensus substrate, bis-(N-benzyloxycarbonyl-L-aspartyl-L-glutamyl-L-valyl-aspartic acid amide) conjugated to rhodamine 110 (Z-DEVD-R110) on its carboxyl-terminal side. Proteolytic cleavage liberates rhodamine 110, unquenching its fluorescence. Fluorescence was measured using excitation and emission wavelengths of 498 and 521 nm, respectively.

Copyright and License information:  ©2021 He, Sehrawat, Verma, Navarro-Corcuera, Sidhu, Mauer, Luo, Katsumi, Chen, Shah, Arab, Cao, Kashkar, Gores, Malhi and Shah.
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