The ORGANELLE-ID-RGB® reagent II is a mixture of cell-permeable red fluorescent lysosomal dye, green fluorescent endoplasmic reticulum 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 Details
| Quantity: | 200μl (for 1000 microscopy assays). |
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| Purity: | ≥93% (HPLC) |
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| Shipping: | Dry Ice |
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| Short Term Storage: | -20°C |
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| Long Term Storage: | -80°C |
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| Handling: | Protect from light. Avoid freeze/thaw cycles. |
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| Technical Info/Product Notes: | Wavelength Maxima:
Lysosomal (Red): Excitation: 568nm Emission: 667nm
Endoplasmic Reticulum (Green): Excitation: 440nm Emission: 565nm
Nuclear (Blue): Excitation: 350nm; Emission: 461nm
The ORGANELLE-ID-RGB® Reagent II 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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| Protocol: | Wide Field Fluorescence/Confocal Microscopy
Reagent Preparation: Mix 2 μl of ORGANELLE-ID-RGB® reagent II in 1 ml of buffer of choice. This volume is sufficient for 10 assays and may be scaled according to need.
Staining Adherent cells:- Grow cells directly onto glass slides or polystyrene tissue culture plates until ~80% confluent via standard tissue culture practices.
- Remove growth media.
- Dispense the freshly diluted staining solution in a volume sufficient for covering the cell monolayer.
- Protect samples from light and incubate for 30 minutes at 37°C.
- Remove the excess staining solution and, if necessary, add a few drops of buffer to prevent the cells from drying out.
- 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:- Grow the cells via standard tissue culture practices.
- Collect about 1 x 105 cells. Centrifuge at 500 x g for 5 minutes. Remove supernatant.
- Re-suspend cells in a volume of the freshly diluted staining solution sufficient for covering the cell pellet.
- Protect the samples from light and incubate for 30 minutes at 37°C.
- Centrifuge at 500 x g for 5 minutes. Remove supernatant.
- Re-suspend the cells in 100 µl buffer.
- Plate 10-15 µL of cells on a glass slide.
- 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).
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| Regulatory Status: | RUO - Research Use Only |
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Figure 1: Live HeLa cells treated with Chloroquine and stained with ORGANELLE-ID-RGB® reagent II showing composite of (A) lysosomes in red, (B) endoplasmic reticula in green, and (C) nuclei in blue.
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General Literature References
Constitutive expression of the machinery for programmed cell death: M. Weil, et al.; J. Cell Biol.
133, 1053 (1996),
Abstract;
Principles and Methods of Toxicology, Third Edition: A.W. Hayes, Ed.; Raven Press 1231 (1994), Book,
