Description
Description: Magic Red Caspase 3 and 7 Assay Kits measure apoptosis-associated DEVDase enzyme activity in living, intact cells. The presence of the active forms of caspases 3 and 7 in apoptotic cells causes the hydrolysis of the two DEVD target sequences from the cresyl violet fluorophore (Magic Red), converting it to the fluorescent form. When the cresyl violet dye is di-substituted with two DEVD caspase-3 /7 target sequences, the reagent is essentially non- fluorescent. Because the Magic Red probe is cell membrane permeant, a cell lysis step is not required to perform the assay. As apoptosis progresses and caspase activity increases, the red fluorescent signal increases. Magic Red (cresyl violet) excites at 540-590 nm (590 nm optimal) and emits at >610nm (630 nm optimal). Detection of DEVD target sequence hydrolysis via the use of the Magic Red fluorogenic substrate assay is a reliable method for monitoring apoptosis induction events and caspase 3&7 activity over time in whole, living cells. Each assay kit includes the Magic Red-(DEVD)2 substrate reagent, Hoechst 33342 stain, and acridine orange stain. This allows cells to be simultaneously evaluated for both the presence of apoptotic nuclear morphology and caspase-3/7 DEVDase activity in fluorescence microscopy analysis techniques. Acridine Orange dye is included to visualize lysosomal organelle structure. Analysis by fluorescence plate reader or flow cytometer may also be performed if the instrument is equipped with a green laser excitation option.
Background: Magic Red Caspase 3 Assay for Apoptosis Detection via Activated Caspase 3 and 7. Caspase-3 is a member of the apoptosis executioner group that also includes caspases -6 and -7 [1]. The inactive zymogen form of caspase-3 is efficiently processed by any of the initiator caspases (caspase-8, caspase-9, and caspase-10). All executioner caspases will target the various cytoskeletal structural proteins as well as PARP and ICAD leading to the morphological and biochemical changes that drive the apoptotic process [2]. Detect Caspase 3 and 7 Activity in Real Time with Magic Red Detection of DEVDase enzyme activity (caspase 3 activity) is a reliable method for assessing apoptotic activity in experimental cell populations [3]. Magic Red Caspase 3 and 7 Assay Kits measure apoptosis-associated DEVDase enzyme activity in living, intact cells via the red fluorescent Magic Red caspase 3 and 7 fluorogenic substrate, MR-(DEVD)2, which is primarily targeted by caspases 3 and 7 of the apoptotic protease cascade. The presence of the active form of caspases 3 and 7 in apoptotic cells causes the hydrolysis of the two DEVD target sequences from the cresyl violet fluorophore (Magic Red), converting it to the fluorescent form. When the cresyl violet dye is di-substituted with two DEVD caspase-3/7 target sequences, the reagent is essentially non-fluorescent. As apoptosis progresses and caspase activity increases, the red fluorescent signal increases. Magic Red (cresyl violet) excites at 540-590 nm (590 nm optimal) and emits at >610nm (630 nm optimal). Because the Magic Red substrate probe is cell membrane permeant, a cell lysis step is not required to perform the assay