Mito-ID
® Green Detection Kit contains a proprietary membrane-permeable mitochondria-selective dye suitable for use with live-, detergent-permeabilized- and aldehyde-fixed-cells. Unlike conventional dyes (such as DiOC6(3), JC-1, rhodamine 123 and tetramethylrhodamine ethyl ester) Mito-ID
® Green dye highlights mitochondria regardless of their energetic state. The dye is compatible with most fluorescence detection systems, including conventional and confocal fluorescence microscopes as well as High Content Screening (HCS) platforms.
The kit is useful for assessing mitochondrial morphology changes, estimating mitochondrial mass and co-localizing RFP-tagged proteins to the mitochondrial compartment.
Figure 1: The Mito-IDâ„¢ Green dye selectively stains mitochondria of living cells (A) and is relatively insensitive to mitochondrial membrane potential uncouplers of phosphorylation, such as CCCP (carbonyl cyanide 3-chlorophenylhydrazone) (B) as well as ion-channel perturbing drugs, such as valinomycin (C).
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Product Specification
| Application: | This kit has been specifically designed for use with RFP-expressing cell lines, as well as cells expressing blue or cyan fluorescent proteins (BFPs, CFPs). |
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| Quality Control: | A sample from each lot of Mito-ID® Green Detection Kit is used to stain HeLa cells, using the procedures described in the user manual. The selectivity of the Mito-ID® Green dye is evident. |
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| Quantity: | 500 assays |
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| Short Term Storage: | -20°C |
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| Long Term Storage: | -80°C |
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| Kit/Set Contains: |
|
Name |
Amount |
|
Mito-ID® Green Detection Reagent |
50μl |
|
Hoechst 33342 Nuclear Stain |
50μl |
|
10x Assay Buffer |
15ml |
|
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| Miscellaneous/General: | This kit has been specifically designed for use with RFP-expressing cell lines, as well as cells expressing blue or cyan fluorescent proteins (BFPs, CFPs). One important application of Mito-ID® Green dye is in fluorescence co-localization imaging with red fluorescent protein (RFP)-tagged proteins. This is a powerful approach for determining the targeting of molecules to intracellular compartments, and for screening of associations and interactions between these molecules. Additionally, the kit is suitable for use with live or post-fixed cells in conjunction with fluorescent probes, such as labeled antibodies, or other fluorescent conjugates displaying similar spectral properties as coumarin, phycoerythrin or cyanine-5. A nuclear counterstain (Hoechst 33342) is provided to highlight this organelle as well. |
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| Background / Technical Information: |
The Mito-ID® Green 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. |
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General Literature References
Photoconversion of Lysotracker Red to a green fluorescent molecule.: E. C. Freundt et al.; Cell. Res.
17, 956 (2007),
Abstract;
Systematic colocalization errors between acridine orange and EGFP in astrocyte vesicular organelles.: F. Nadrigny et al.; Biophys. J.
93, 969 (2007),
Abstract;
Chloromethyl-X-rosamine (MitoTracker Red) photosensitises mitochondria and induces apoptosis in intact human cells.: T. Minamikawa et al.; J. Cell. Sci
112, 2419 (1999),
Abstract;
Chloromethyltetramethylrosamine (Mitotracker Orange) induces the mitochondrial permeability transition and inhibits respiratory complex I. Implications for the mechanism of cytochrome c release. : L. Scorrano et al.; J. Biol. Chem.
274, 24657 (1999),
Abstract;