Mito-ID^® Membrane potential detection kit for microscopy and flow cytometry



ENZ-51018-K100	1 Kit	407.00 USD

The only assay that monitors energetic status using a simple mix-and-read, no-wash protocol

Dual-emission dye fluoresces either green or orange, depending upon mitochondrial membrane potential status
Highly sensitive detection of mitochondrial membrane potential decline
Aids in the drug research process in preclinical drug safety assessment (ADME-Tox)
Suitable for chemical/environmental toxicity screening and characterization
True mix-and-read homogeneous assay for live cells
Optimized for fluorescence microscopy and flow cytometry

Cell-based assays for evaluating the functional status of mitochondria are emerging as useful tools for elucidating the role of mitochondrial activity in drug-induced toxicity, the apoptosis cascade and other cellular and biochemical processes. The loss of the mitochondrial membrane potential (MMP) is often associated with early stages of apoptosis. The collapse of MMP coincides with the opening of the mitochondrial permeability transition pores, leading to the release of cytochrome C into the cytosol, which in turn triggers other downstream events in the apoptotic cascade.

Mito-ID^® Membrane Potential Detection Kit measures mitochondrial membrane potential with a cationic dye that fluoresces either green or orange depending upon membrane potential status.

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Figure 2: Using a conventional fluorescence microplate reader, Mito-ID™ Membrane Potential dye was shown to decrease as a function of CCCP concentration (decrease in orange signal). Mito-ID™ Membrane Potential dye is at least 10-fold more sensitive to mitochondrial potential loss than the commonly used dye, JC-1. The high Z-factor (≥0.9) obtained using the Mito-ID™ Membrane Potential dye arises from the no-wash protocol.

Figure 3: Treatment of HeLa Cells with K+ Valinomycin. Using a conventional fluorescence microplate reader, mitochondria membrane potential was shown to decrease as a function of valinomycin concentration. The high Z-factor (≥0.75) obtained using the Mito-ID™ Membrane Potential dye demonstrates excellent signal-to-noise ratios.

Figure 4: Treatment of HeLa Cells with 2,4 Dinitrophenol. Using a conventional fluorescence microplate reader, mitochondrial membrane potential was shown to decrease as a function of 2,4 Dinitrophenol concentration. The high Z-factor (≥0.7) obtained using the Enzo Mito-ID™ Membrane Potential dye demonstrates excellent signal-to-noise ratios.

Figure 5: Flow Cytometric Analysis of Control and Treated Cells. Jurkat cells were untreated (left) and were treated with 1 μM CCCP for 15 mins (right). Cells were then stained with Enzo Mito-ID™ Membrane Potential Dye, and run on a FACS Calibur instrument.

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Product Specification

Quantity:	100 assays

Quality Control:	1. % purity of Mito-ID^® MP Detection Reagent by TLC ≥ 95% 2. A sample kit from each lot of Mito-ID^® Membrane Potential Detection Kit for fluorescence microscopy and flow cytometry is assayed using a microplate-based assay. The Z’-factor from CCCP-treated cells is greater than 0.6.

Kit/Set Contains:	Mito-ID^® MP Detection Reagent, 500μl Necrosis Detection Reagent, 600μl CCCP Control, 100μl 10X Assay Buffer 1: 25ml 50X Assay Buffer 2: 5ml

Application:	The Mito-ID^® Membrane Potential Detection Kit has been optimized for measurement of mitochondria membrane potential (MMP) and cell viability by conventional fluorescence microscopy and flow cytometry.

Short Term Storage:	-20°C

Long Term Storage:	-80°C

Handling:	Avoid freeze/thaw cycles. Protect from light.

Background / Technical Information:	The Mito-ID^® Membrane Potential Detection Kit is a member of the CELLestial^® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications. CELLestial^® reagents and kits are optimal for use in demanding cell analysis applications involving confocal microscopy, flow cytometry, microplate readers and HCS/HTS, where consistency and reproducibility are required.

General Literature References

High throughput fluorescence assays for the measurement of mitochondrial activity in intact human neuroblastoma cells: A.J. Woollacott & P.B. Simpson; J. Biomol. Screen 6, 413 (2001), Abstract;

A New Method for the Cytofluorometric Analysis of Mitochondrial Membrane Potential Using the J-Aggregate Forming Lipophilic Cation 5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine Iodide (JC-1): A. Cossarizza, et al.; Biochem. Biophys. Res. Commun. 197, 40 (1993), Abstract;

Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate forming lipophilic cation JC-1: S.T. Smiley, et al.; PNAS 88, 3671 (1991), Full Text