Organelle-ID™ RGB reagent I

Mixture of fluorescent dyes for detection of lysosomes, mitochondria and nucleic acids



ENZ-53007-C200	200 µl	319.00 USD

The Organelle-ID™ RGB reagent I is a mixture of cell-permeable red fluorescent lysosomal dye, green fluorescent mitochondrial dye and blue fluorescent nucleic acid dye. The staining pattern arising from the combination of these three dyes permits visualization of the target organelles by fluorescence/confocal microscopy. The reagent, supplied as a 500X solution, is sufficient for 1000 microscopy assays. The single-tube format makes this multi-organelle stain reagent easy to use.

Product Specification

Quantity:	200μl (for 1000 microscopy assays).

Purity:	≥93% (HPLC)

Short Term Storage:	-20°C

Long Term Storage:	-80°C

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

Background / Technical Information:	Wavelength Maxima: Lysosomal (Red): Excitation: 568nm Emission: 667nm Mitochondrial (Green): Excitation: 460nm Emission: 560nm Nuclear (Blue): Excitation: 350nm Emission: 461nm The Organelle-ID™ RGB Reagent I 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.

Protocol:	Wide Field Fluorescence/Confocal Microscopy Reagent Preparation: Mix 2 μL of Organelle-ID™ RGB reagent I in 1 mL of buffer of choice. This volume is sufficient for 10 assays and may be scaled according to need. Staining Adherent cells: 1. Grow cells directly onto glass slides or polystyrene tissue culture plates until ~80% confluent via standard tissue culture practices. 2. Remove growth media. 3. Dispense the freshly diluted staining solution in a volume sufficient for covering the cell monolayer. 4. Protect samples from light and incubate for 30 minutes at 37°C. 5. Remove the excess staining solution and, if necessary, add a few drops of buffer to prevent the cells from drying out. 6. Cover cells with a glass cover slip and observe under a fluorescence/confocal microscope with a filter set for DAPI (Ex/Em: 350/470nm), Texas Red (Ex/Em: 540/605 nm) and GFP/FITC (Ex/Em: 488/514 nm). Staining Non-Adherent cells: 1. Grow the cells via standard tissue culture practices. 2. Collect about 1 x 10⁵ cells. Centrifuge at 500 x g for 5 minutes. Remove supernatant. 3. Re-suspend cells in a volume of the freshly diluted staining solution sufficient for covering the cell pellet. 4. Protect the samples from light and incubate for 30 minutes at 37°C. 5. Centrifuge at 500 x g for 5 minutes. Remove supernatant. 6. Re-suspend the cells in 100 µL buffer. 7. Plate 10-15 µL of cells on a glass slide. 8. Cover cells with a glass cover slip and observe under a fluorescence/confocal microscope with a filter set for DAPI (Ex/Em: 350/470nm), Texas Red (Ex/Em: 540/605 nm) and GFP/FITC (Ex/Em: 488/514 nm).

Figure 1: Live cells stained with Organelle-ID™ RGB reagent I showing composite of (A) nuclei in blue, (B) mitochondria in green, and (C) lysosomes in red.

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