MITOCHONDRIAL MEMBRANE POTENTIAL CYTOCHROME C RELEASE APOPTOSIS DETECTION KITS FOR IN VITRO USE JC-1 JC1 TMRE TMRM

MitoPT™ Mitochondrial Membrane Permeability Transition Kits
ICT now has 3 kits to easily assess changes in mitochondrial membrane potential. Changes in mitochondrial membrane potential can correlate with cytochrome c release and the initiation of apoptosis. 

Jurkat cells were stained with ICT’s MitoPT™-JC1 dye. Healthy cells fluoresce red, dying cells fluoresce green.

NEW!     1. MitoPT™-TMRE red fluorescence
NEW!     2. MitoPT™-TMRM red fluorescence 
                3. MitoPT™-JC1 green and red fluorescence

• Easy Just add directly to the media; healthy cells will fluoresce orange/red. 
• Fast Typical incubation is 15-20 minutes.
• Accurate Orange/red signal is reduced in dying cells.
• Sensitive Easily distinguish positive from negative populations.
• Quantitative Analyze with a flow cytometer, fluorescence plate reader, or fluorescence microscope.
• Whole cell analysis The reagents are cell-permeant so you do not have to lyse the cells. 

Order now!  1-800-829-3194.

Red MitoPT™-TMRE

 MitoPT™-TMRE kits contain: 
1. MitoPT™-TMRE reagent 
2. CCCP, a Depolarizing reagent
3. Assay buffer

Manual 

Order now!  1-800-829-3194.

MitoPT-TMRE is
easy
fast
sensitive.

Try it today
1-800-829-3194
Order

Red MitoPT™-TMRM

 MitoPT™-TMRM kits contain: 
1. MitoPT™-TMRM reagent 
2. CCCP, a Depolarizing reagent
3. Assay buffer

Manual 

Order now!  1-800-829-3194.

MitoPT-TMRM is
easy
fast
sensitive.

Try it today
1-800-829-3194
Order

IMPROVED!!  Green & Red MitoPT™-JC1

Healthy cells fluoresce red, dying cells fluoresce green. Jurkat cells were stained with ICT’s MitoPT™-JC1 dye and analyzed with a fluorescence microscope containing a long band path filter (excitation at 490 nm and emission >510 nm). In non-apoptotic healthy cells (2 cells on top), the reagent aggregates inside intact mitochondria and fluoresces red. As the mitochondrial membrane potential drops and cells enter apoptosis, the dye disperses throughout the cell. The reagent then assumes a monomeric form and fluoresces green (3 cells on bottom).

Manual 

      ICT’s original MitoPT™-JC1 Mitochondrial Permeability Transition kit has been improved! You can test a lot more samples, and it includes CCCP, a depolarizing agent. MitoPT-JC1 can be used to quantitate apoptosis and mitochondrial functionality - non-apoptotic cells with healthy mitochondria appear red, apoptotic cells appear green.  
     ICT’s unique MitoPT™-JC1 reagent is based on a fluorescent cationic dye, commonly known as JC-1. Just culture your cells and add MitoPT™-JC1 to the media. MitoPT™-JC1 easily penetrates cells and healthy mitochondria where it aggregates and fluoresces red. As the mitochondrial membrane potential collapses, MitoPT™-JC1 is diffused throughout the cell. Once dispersed, the reagent assumes a monomeric form and fluoresces green. This can be read with a fluorescence plate reader, flow cytometer, or fluorescence microscope. MitoPT™-JC1 excites at 488nm, and emits at >590nm. 

Sample protocol for suspension cells (read manual):

1. Culture cells up to 1 x 10⁶ cells/mL. 
2. Induce apoptosis following your protocol. 
3. Create controls using CCCP or other agent. 
4. Reconstitute MitoPT JC1 with DMSO to form the stock concentrate (can be frozen for future use). 
5. Dilute stock concentrate with 1X Assay Buffer to form the working solution. 
6. Add the working solution directly to your cell culture for labeling. 
7. Incubate 15 minutes -1 hour. 
8. Wash cells twice.
9. Analyze data using a fluorescence microscope, plate reader, or flow cytometer.

MitoPT-JC1  kit, catalog. #911 Kit contains: MitoPT™-JC1 reagent, CCCP, and assay buffer.

Order now!  1-800-829-3194.

MitoPT™-JC1 is
easy
fast
sensitive
quantitative

Try it today
1-800-829-3194
Order

Read cells with a fluorescence microscope, plate reader, or flow cytometer.

Using a flow cytometer to analyze cells labeled with MitoPT™-JC1, the instrument will measure apoptosis by monitoring the amount of red fluorescence in each region. Healthy cells with intact mitochondria fluoresce red due to aggregated MitoPT™-JC1 and appear in R2. As the mitochondrial membrane permeability collapses and cells enter apoptosis, MitoPT™-JC1 is dispersed throughout the cell where it converts to a monomeric form and fluoresces green. The amount of red fluorescence drops as these cells enter R3. In this example, Jurkat cells were either treated with DMSO (negative, non-induced cells) or with staurosporine (apoptotic, induced cells) for 4 hours and then labeled with MitoPT™-JC1 for 15 minutes.

Read all references (pdf).