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Calpain activity measurement

JZ Ji-Na Zheng
YL Yang Li
YY Yue-Mei Yan
YY Yong Yu
WS Wen-Qi Shao
QW Qiang Wang
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Calpain activity kit (Raybiotech, USA) was utilized to measure calpain activities in serum or plasma. Eighty-five microliters of serum was diluted in 10 μL of 10X calpain reaction buffer and 5 μL of calpain substrate Ac-LLY-AFC with or without 100 μM calpeptin (Abmole, USA). Free AFC was quantified using a fluorometer (excitation λ 400 nm, emission λ 505 nm) after incubating at 37 °C for 1 h in the dark. The difference of calpain activity was determined by comparing the relative fluorescent unit (RFU) of samples with and without calpeptin. The calpain activity was expressed as RFU per microliter serum of each sample.

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