The activity of caspase-3, -8 and -10 was measured using caspase colorimetric protease assay kits (Millipore, Billerica, MA, USA) according to the manufacturer’s instructions. In brief, the A549 cells were treated with DDP and GEN alone or in combination for 24 h. Following treatment, the cells were washed twice with ice-cold PBS and harvested by centrifugation at 1,000 × g for 10 min. The cell pellets were then lysed in 150 μl buffer. Protein concentrations in the lysate were determined using the Lowry method (16 (link)). An 80-μl aliquot of the lysate was incubated with 10 μl of substrate for each caspase at 37°C for 2 h. The samples were analyzed at 405 nm in a microplate reader (Thermo Fisher Scientific Inc.). The relative caspase activity of the control group was set as 100.
Caspase colorimetric protease assay kits
Manufactured by Merck Group
Sourced in United States
The Caspase colorimetric protease assay kits are laboratory equipment designed to detect and quantify the activity of caspase enzymes. These kits provide a simple and reliable method for measuring caspase activity in cell lysates, purified enzyme preparations, or other biological samples.
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Lab products found in correlation
2 protocols using caspase colorimetric protease assay kits
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Caspase Protease Activity Quantification
2
Evaluating Caspase Enzyme Activity
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To evaluate caspase activity, caspase colorimetric protease assay kits (Millipore, Billerica, MA) were used. In brief, harvested cell pellets were lysed in the lysis buffer, and supernatants were collected. Equal amounts of protein were incubated with reaction buffer and colorimetric substrate, acetyl (Ac)-Ile-Glu-Thr-Asp (IETD) p-nitroaniline (pNA) for caspase-8, Ac-Leu-Glu-His-Asp (LEHD)-pNA for caspase-9, and Ac-Asp-Glu-Val-Asp (DEVD)-pNA for caspase-3, respectively, at 37℃ for 2 h in the dark. Thereafter, the reactions were assessed for changes in absorbance at 405 nm using a microplate reader. In the caspase-3 inhibitor assay, cells were pretreated with a caspase-3 inhibitor (50 µM zDEVD-fmk) for 1 h prior to the addition of HEABG [25 ].
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