MitoSpy™ NIR DiIC1(5)

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Other Names
Mitochondrial labeling, DiIC1(5)
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MitoSpy_NIR_1_ICC_062118
HeLa cells were stained with 0.25µM CytoPhase™ Violet dye (red) for 60 minutes at 37°C. Then 20 nM of MitoSpy™ NIR DiIC1(5) (green) was added for an additional 30 minutes at 37°C. The Z-stack images were captured on an LSM 880 with Airyscan using a 63x Oil objective. The files were Airyscan processed and a Maximum Intensity projection was created using Zen software; all courtesy of Biophotonics Core Facility at Salk Institute.
  • MitoSpy_NIR_1_ICC_062118
    HeLa cells were stained with 0.25µM CytoPhase™ Violet dye (red) for 60 minutes at 37°C. Then 20 nM of MitoSpy™ NIR DiIC1(5) (green) was added for an additional 30 minutes at 37°C. The Z-stack images were captured on an LSM 880 with Airyscan using a 63x Oil objective. The files were Airyscan processed and a Maximum Intensity projection was created using Zen software; all courtesy of Biophotonics Core Facility at Salk Institute.
  • MitoSpy_NIR_2_FC_062118
    Human T-cell leukemia cell line, Jurkat, was treated (left) or un-treated (right) with 1.0 µg/ml of LEAF™ purified anti-human CD95. At the end of incubation, cells were washed twice, (once with PBS then second time with Annexin V Binding Buffer), then were stained with 5 nM of MitoSpy™ NIR DilC1(5), Brilliant Violet 421™ Annexin V, and Helix NP™ Green (Cat. No. 425303 at 1.25 nM) for 15 minutes at 37°C in Annexin V Binding Buffer (Cat. No. 422201). After incubation, cells were washed and resuspended with Annexin V Binding Buffer. Shown gated on live cells.
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424807 200 tests $50
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Description

MitoSpy™ mitochondrial localization probes are cell-permeant, fluorogenic chemical reagents that are used for labeling mitochondria in live cells. MitoSpy™ NIR DilC1(5) localizes to the mitochondria based on its membrane potential and is useful to indicate cell health as well as for localization. Only useful on live cells, the use of fixatives is not recommended prior to imaging or flow cytometry.

Product Details
Technical data sheet

Product Details

Molecular Mass
510.45 MW
Preparation
The stock solution for MitoSpy™ NIR DilC1(5) is prepared by dissolving the lyophilized probe in dimethyl sulfoxide (DMSO) to make a final concentration of 10uM by adding 500uL of DMSO to each vial.
Storage & Handling
Store MitoSpy™ NIR DilC1(5) at -20°C.
Application

ICC/Live cell imaging - Quality tested
FC - Validated

Recommended Usage

Each lot of this reagent is quality control tested by immunocytochemistry staining. For immunocytochemistry microscopy, a concentration range of 1 to 50 nM is recommended. For flow cytometric staining, a concentration range of 1 to 50 nM is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

MitoSpy™ NIR DilC1(5) is excited at 638 nm and emits at 658 nm.

Protocol for Live Cell Microscopy

  1. Reconstitute MitoSpy™ NIR DilC1(5) to a 10uM concentration with DMSO by adding 500uL to an individual vial of lyophilized material. Protect the stock solution from light and keep frozen for storage.
  2. Prepare the working solution for MitoSpy™ NIR DilC1(5) in 37°C pre-warmed culture media, which will vary by cell line and type of imaging required. When labeling mitochondria for live cell imaging a concentration of 1-50 nM is recommended.
  3. Grow cells to a desired confluency and wash once with pre-warmed 1X PBS.
  4. Add the working solution of MitoSpy™ NIR DilC1(5) to the live cells and incubate at 37°C for 20-30 minutes.
  5. Wash the cells twice with warm 1XPBS or culture media.
  6. Cells are now ready to be imaged live with a fluorescence microscope. MitoSpy™ NIR DilC1(5) is poorly retained with PFA fixation.

Antigen Details

Distribution

Mitochondria

Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Mitochondrial Function, Neuroscience
Molecular Family
Mitochondrial Markers
Gene ID
NA

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Go To Top Version: 1    Revision Date: 06/27/2018

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

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