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Antibodies

JC-1 Fluorescent Mitochondrial Membrane Depolarization Assay

Product Sizes
100 tests
924-100TESTS
About this Product
SKU:
924
Application:
Flow Cytometry
CE/IVD:
RUO
Extra Details:
JC-1 accumulates and fluoresces orange in polarized mitochondria. Following membrane depolarization, JC-1 disperses within the cell and fluoresces green. Analyze using a flow cytometer, fluorescence plate reader, or fluorescence microscopy., ImmunoChemistry Technologies
Immunogen:
Mitochondrial depolarization
Physical State:
Cell Culture
Purification:
Healthy cells fluoresce red, dying cells fluoresce green. Jurkat cells were stained with ICT's MitoPT JC-1 dye and analyzed with a fluorescence microscope containing a long bandpass filter (excitation
Shipping Conditions:
Blue Ice
Storage Conditions:
-20[o]C
Type:
Antibody: Stains & Enhancers
Figure 1. Jurkat cells were treated with DMSO (negative control) or 1 uM staurosporine for 2 hours to induce apoptosis. Cells were then stained with JC-1 dye. Normal healthy cells (two cells, upper and lower left) concentrate JC-1 in mitochondria and fluoresce bright orange. Apoptotic cells (three cells, right), with mitochondria of various stages of permeability, exhibit a reduced orange fluorescence and increased green fluorescence, as JC-1 disperses through the cells. (Dr. Brian Lee, ICT).
Figure 2. Jurkat and HL-60 cells were exposed to DMSO (negative control) (dark orange) or 50 uM CCCP depolarizing agent (light orange) and then incubated with JC-1 for 20 minutes at 37°C, and washed. Aliquots were analyzed in triplicate in a fluorescence plate reader set at 488 nm excitation and 590 nm emission filter settings. Healthy cells exhibited a high level of orange fluorescence; the CCCP-depolarized cells had reduced orange fluorescence as the dye dispersed. (Ms. Tracy Hanson, ICT)
Figure 3. Jurkat cells were treated with 1 uM staurosporine, an apoptosis-inducing agent (black), or DMSO (negative control) (red), stained with JC-1, and analyzed by flow cytometer. Apoptotic cells (black) exhibit much less fluorescence compared to healthy cells (right) which concentrate JC-1 in their polarized mitochondria. Almost all cells treated with staurosporine exhibit apoptosis-associated mitochondrial depolarization (Ms. Tracy Hanson, ICT, 13A54, 050113).
Figure 4. Jurkat cells were treated with either DMSO (Negative Control) or staurosporine (Positive Control) for 3 hours at 37°C and labeled with JC-1 for 15 minutes. Cells were analyzed with a FACS Caliber Becton Dickinson Flow Cytometer. Collapse of the mitochondrial membrane potential gradient is indicated by an increase in the number of cells falling from P2 (Green) into P3 (Pink) following induction of apoptosis (Dr. Brian Lee, ICT).