caspase 3 substrate  (Biotium)

Journal: bioRxiv

Article Title: Selective granzyme B replenishment dominates the molecular switch for sustained cytotoxicity in effector CD8 + T lymphocytes

doi: 10.64898/2026.10.09.758067

Figure Lengend Snippet: Target cells were preloaded with a fluorogenic GzmB substrate (GranToxiLux) and conjugated with fresh or pre-exposed CTLs. (A) Representative time-lapse images showing GranToxiLux cleavage in target cells (green and pseudocolor) following interaction with fresh or pre-exposed CTLs; (B) Flow cytometric quantification of GranToxiLux cleavage in target cells at 30 min and 1 h after conjugation. Left, representative histogram; right, pooled quantification from n = 3 independent experiments. (C) Representative flow cytometry quantification of cleaved caspase-3 in target cells following conjugation with CTLs (same experimental conditions as in B). Left, representative histogram; right, pooled quantification from at least three independent experiments. (D) Representative snapshots from live-cell imaging showing PI uptake (red) in target cells as a marker of membrane permeabilization and cell death. Top, non-exposed/fresh (NE) CTLs; bottom, pre-exposed (PE) CTLs. White arrowheads indicate T cells. (E) Time to PI entry for target cells killed by non-exposed/fresh (NE) or pre-exposed (PE) CTLs. Data are shown as mean +/-s.e.m. Statistical significance was assessed using unpaired t-test. ns, not significant, *P < 0.05; ***P < 0.001.

Article Snippet: Alternatively, target cells were pre-loaded with a caspase-3 substrate (NucView 488, Biotium) at 5 μM in complete medium for 30 min at 37 °C, washed twice, and conjugated with CTLs for the indicated time points.

Techniques: Conjugation Assay, Flow Cytometry, Live Cell Imaging, Marker, Membrane

Journal: Frontiers in Pharmacology

Article Title: A novel quinine derivative as a PIM-1 kinase inhibitor induces apoptosis via mitochondrial depolarization with selective cytotoxicity in acute lymphoblastic leukemia cells

doi: 10.3389/fphar.2026.1799674

Figure Lengend Snippet: A significant activation of caspase-3/7 was observed after treating CEM cells for 5 h with compound 22 .

Article Snippet: Caspase-3/7 activation was assessed in CEM cells using the NucView 488 caspase-3 fluorogenic substrate (Biotium, #10402, Fremont, USA).

Techniques: Activation Assay

Journal: bioRxiv

Article Title: Synergistic induction of a lethal Autosis-to-Apoptosis switch by Phytocannabinoids and beta-Caryophyllene in Triple-Negative Breast Cancer Cells

doi: 10.64898/2026.04.05.716056

Figure Lengend Snippet: (A) MDA-MB-231 cells were treated with vehicle (0.3% DMSO - Control) or combinations C1 to C3 as described in in the presence of SYTOX Green and SPY-DNA. Automated live-cell imaging was performed for 72h and representative composite images combining the phase contrast, green (SYTOX Green, dead cells), and NiR (SPY650-DNA, nuclei) fluorescence channels obtained at different time points are displayed. Dead cells (SPY + /SYTOX Green + nuclei) are indicated by white arrows. Insets show higher magnifications of specific elements. Blue boxes highlight retracted cells exhibiting cytoplasmic vacuoles, yellow boxes indicate cells displaying extensive cytoplasmic blebbing, and orange boxes highlight the formation of large cytoplasmic blisters (orange arrows) characteristic of secondary necrosis. Scale bar corresponds to 200 µm. See Videos S1-4 (Supplementary Material) for the complete 72 h time-lapse sequences. (B) MDA-MB-231 cells were treated with combinations C1 in the presence of SYTOX Orange and NucView 488 Caspase-3 substrate. Automated live-cell imaging was performed and representative phase contrast and composite images combining the green (NucView, active caspase-3) and orange (SYTOX Orange, dead cells) fluorescence channels obtained at different time points are displayed. Retracted cells are indicated by white arrow heads.

Article Snippet: To evaluate apoptosis, the NucView® 488 Caspase-3 enzyme substrate (0.5 μM; Biotium, USA) was used, with staurosporine (0.5 μM) as a specific positive control for caspase activation.

Techniques: Control, Live Cell Imaging, Fluorescence

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Thermal Sensitizers That Inhibit Heat-Induced SAFB Granule Formation

doi: 10.1021/acs.jmedchem.5c03361

Figure Lengend Snippet: Apoptosis induction efficiency in the presence of compounds. (A) Schematic representation of cell treatment with the compounds used to detect the apoptosis induction efficiency. Apoptosis efficiencies were detected using NucView488 (green) and propidium iodide (PI) (red) in HeLa (B), H1299 (C), AsPC1 (D), HepG2 (E), and HEK293T (F) cell lines treated with 5 μM compounds or 50 μM 5-fluorouracil (5-FU) at 37 or 43 °C. Nonapoptotic cell death was detected using PI. Blue bars, NucView- and PI-stained cells; red bars, NucView-stained cells; and green bars, PI-stained cells. Data were presented as the mean ± standard error of six independent experiments. Over 200 cells were counted in each experiment. Cisplatin (5 μM) and 5-FU (50 μM) were used as positive controls.

Article Snippet: Apoptosis was detected using the NucView488 caspase-3 substrate (Biotium, Fremont, CA) and PI (Molecular Probes, Eugene, OR).

Techniques: Staining