Devd sequence

Manufactured by Promega

The DEVD sequence is a short amino acid sequence that can be used as a substrate for the detection and measurement of caspase-3 and caspase-7 activity in biological samples. It provides a reliable and specific means to assess these key enzymes involved in apoptosis, or programmed cell death.

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Lab products found in correlation

Calcein am, by AnaSpec (2 mentions) Propidium iodide, by Roche (2 mentions)

2 protocols using devd sequence

Caspase-3/7 and Viability Assay

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SKOV-3 or MEF cell lines were pretreated with 10 μm LPA (Avanti) for 1 h in 0.1% fatty acid-free BSA-containing medium, followed by incubation with 50 μm cisplatin for 20 h (SKOV-3) or 2 μm adriamycin or 8 h (MEFs). Caspase-3/7 activity was determined by cleavage of the luminogenic substrate containing the DEVD sequence (Promega) and was normalized to protein concentrations. Alternatively, cell viability was determined by calcein/propidium iodide double fluorescence staining of the live/dead cells. After treatment was complete, cells were incubated with 0.5 μm calcein-AM (AnaSpec, Fremont, CA, USA) and 0.1 μg ml⁻¹ propidium iodide (Roche, Indianapolis, IN, USA) for 10 min at room temperature. Images of green fluorescent calcein-positive live cells and red fluorescent propidium iodide-positive dead cells were acquired under fluorescence microscope using ×20 long-ranged objectives. Totally 500–1000 cells per sample were counted to determine the percentage of live and dead cells.

Lin F.T., Lin V.Y., Lin V.T, & Lin W.C. (2016). TRIP6 antagonizes the recruitment of A20 and CYLD to TRAF6 to promote the LPA2 receptor-mediated TRAF6 activation. Cell Discovery, 2, 15048-.

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Assessing Cytotoxicity and Apoptosis

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SKOV-3 or MEF cell lines were pretreated with 10 μM LPA (Avanti) for 1 h in 0.1% fatty acid-free BSA-containing medium, followed by incubation with 50 μM cisplatin for 20 h (SKOV-3) or 2 μM adriamycin or 8 h (MEFs). Caspase-3/7 activity was determined by cleavage of the luminogenic substrate containing the DEVD sequence (Promega) and was normalized to protein concentrations. Alternatively, cell viability was determined by calcein/propidium iodide double fluorescence staining of the live/dead cells. After treatment was complete, cells were incubated with 0.5 μM calcein-AM (AnaSpec, Fremont, CA, USA) and 0.1 μg ml⁻¹ propidium iodide (Roche, Indianapolis, IN, USA) for 10 min at room temperature. Images of green fluorescent calcein-positive live cells and red fluorescent propidium iodide-positive dead cells were acquired under fluorescence microscope using × 20 long-ranged objectives. Totally 500–1000 cells per sample were counted to determine the percentage of live and dead cells.

Lin F.T., Lin V.Y., Lin V.T, & Lin W.C. (2016). TRIP6 antagonizes the recruitment of A20 and CYLD to TRAF6 to promote the LPA2 receptor-mediated TRAF6 activation. Cell Discovery, 2, 15048-.

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