Mito-ID^® Membrane potential cytotoxicity kit for microplates



ENZ-51019-KP002	1 Kit	209.00 USD

Enzo Life Sciences’ Mito-ID^® Membrane Potential Cytoxicity Kit for microplates measures mitochondrial membrane potential with a cationic dye that fluoresces either green or orange depending upon mitochondrial membrane potential status. In energized cells, the Mito-ID^® MP Detection reagent exists as a green-fluorescent monomer in the cytosol and also accumulates as orange-fluorescent aggregates in the mitochondria. However, in cells with compromised mitochondrial membrane potential, the Mito-ID^® MP Detection Reagent dye exists primarily as green-fluorescent monomers throughout the cytosol and no longer exhibits orange fluorescence in the mitochondria.

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Product Specification

Quantity:	For 2 x 96-well microplates

Quality Control:	A sample kit from each lot of Mito-ID^® Membrane Potential Cytotoxicity Kit is assayed using the microplate-based assay described in the manual. The Z’-factor from CCCP-treated cells is greater than 0.6.

Kit/Set Contains:	Mito-ID™ MP Detection Reagent, 200μl CCCP Control, 100μl 10X Assay Buffer 1: 2.5ml 50X Assay Buffer 2: 0.5ml

Application:	Mito-ID^® Membrane Potential Cytotoxicity Kit enables monitoring of mitochondrial potential changes using a simple fluorescence microplate reader.

Shipping:	SHIPPED ON DRY ICE

Short Term Storage:	-20°C

Long Term Storage:	-80°C

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

Background / Technical Information:	For the Manual please click here. The Mito-ID^® Membrane Potential Cytotoxicity 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.

Figure 1: The mitochondria of HeLa cells were stained with Mito-ID™ Membrane Potential reagent, and visualized by epifluorescence microscopy. Orange fluorescent aggregates are localized in the mitochondria (Orange channel), while green fluorescent monomers mainly stain the cytosol (FITC channel).

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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General Literature References

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

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

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