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JC-1 Mitochondrial Membrane Potential Assay Kit

Cat. No. M10454
JC-1 Mitochondrial Membrane Potential Assay Kit Structure
Size Price Availability Quantity
100Tests USD 120  USD120 In stock
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Quality Control
Biological Activity

JC-1 Mitochondrial Membrane Potential Assay Kit is a kit that uses JC-1 as a fluorescent probe to quickly and sensitively detect the transmembrane potential difference of tissues, cells or purified mitochondria. It can be used for early apoptosis detection.

Components Size
M9724 JC-1 3 mM in DMSO * 600 uL
M6567 CCCP 50 mM in DMSO * 600 uL

 

General Protocol

a. Culture cells in 6-, 12- , 24-, or 96-well plates at a density of 5× 105 cells/mL. Incubate the cells according to your normal protocol.

b. For the control tube, allow the vial of CCCP has come to room temperature, add 1 μL of CCCP (50 mM). Incubate cells at 37°C for 5 minutes.

c. Add 5 μL JC-1 (3 mM) per well to make the final concentration at 15 μM. Incubate cells at 37°C, 5% CO2, for 15-20 minutes. 

d. After incubation, centrifuge cells for 3-4 minutes at 500×g at 4°C, carefully aspirate the supernant.

e. Wash cells twice with PBS: add 2 mL PBS to suspend cells and vortex to mix thoroughly. Centrifuge cells for 3-4 minutes at 500×g at 4°C, carefully aspirate the supernant.

f. Add 500 μL PBS to suspend cells. Analyze sample on a flow cytometer, fluorescence microscopy, or fluorescence microplate reader. (Green fluorescence: Ex/Em=510/527 nm; Red fluorescence: Ex/Em=585/590 nm)


Stored at -20°C protecting from light, and is stable for up to 12 months. Avoid repetitive freeze-thaw cycles while using. For immediate use, components may be stored at 2-8°C.


Precautions 

1. A ‘Test’ simply refers to a single assay well. 100 tests based on labeling volumes of 1.0 mL. 

2. JC-1 is light sensitive, we recommend that you make small aliquots and store them at -20℃ with light protected.

 

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)
Species Mouse Rat Rabbit Guinea pig Hamster Dog
Weight (kg) 0.02 0.15 1.8 0.4 0.08 10
Body Surface Area (m2) 0.007 0.025 0.15 0.05 0.02 0.5
Km factor 3 6 12 8 5 20
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

Chemical Information
Molecular Weight
Formula
CAS Number
Purity
Solubility
Storage -20°C, protect from light, sealed
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  Catalog
Abmole Inhibitor Catalog 2017




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