Cell Navigator™ Live Cell RNA Imaging Kit *Green Fluorescence*

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Fluorescence images of RNA staining in HeLa cells. (A) Live cells were stained using Cell Navigator™ Live Cell RNA Imaging Kit (Green, Cat#22630) and counter-stained with Hoechst 33342 (Blue, Cat#17530). (B) Cells fixed in methanol were stained using the same kit. (C) After staining, fixed HeLa cells were incubated with 0.5 mg/mL RNase at 37 ºC for 1 hour. Image of RNase digest test indicates the high selectivity of StrandBrite™ RNA Green. The green fluorescence signal were measured using a fluorescence microscope with a FITC filter.
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Unit Size: Cat No: Price (USD): Qty:
100 Tests 22630 $295


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Additional Ordering Information
Telephone: 1-800-990-8053
Fax: 1-408-733-1304
Email: sales@aatbio.com
International: See distributors





Overview

Ex/Em (nm)503/511
Storage Freeze (<-15 °C)
Minimize light exposure
InstrumentsFluorescence microscope
Category Cell Biology
Labeling Cells
Related Fluorescence Imaging
Detecting and imaging RNA molecules in living cells is extremely important for a wide variety of molecular biology procedures including physical transportation, interpretation of genetic information, regulation of gene expression and some essential bio-catalytic roles. The major challenge to stain RNA in living cells is the interferences caused by DNA. In order to address the difficulty, a novel green fluorogenic dye was developed as a RNA-selective probe. AAT Bioquest's Cell Navigator™ Live Cell RNA Imaging Kit includes StrandBrite™ RNA Green as it specifically binds RNA in cells. Compared to commercial SYTO® RNA Select dye for RNA staining in vivo, StrandBrite™ RNA Green shows brighter signal and much better selectivity to RNA. In addition, this kit can stain RNA in both living cells and fixed cells.




Protocol


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This protocol only provides a guideline, and should be modified according to your specific needs.
At a glance

Protocol summary

  1. Prepare cells
  2. Add StrandBrite™ RNA Green working solution
  3. Incubate for 30 - 60 minutes
  4. Analyze the cells under fluorescence microscope at Ex/Em = 490/520 nm (FITC filter set)

Important notes
Thaw all the components at room temperature before starting the experiment.

Key parameters
Instrument:Fluorescence microscope
Excitation:490 nm
Emission:520 nm
Instrument specification(s):FITC filter set
Recommended plate:Black wall/clear bottom

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

Sample experimental protocol
  1. Culture cells to a density optimum for imaging according to your specific induction protocol (about 1 - 2 × 104 cells/well/96-well plate).

  2. For living cells: Incubate cells with StrandBrite™ RNA Green (Component A) diluted 400X in medium or live cell staining buffer (Component B) at room temperature for 30 - 60 minutes (100 µL/well). Note: 25 µL of StrandBrite™ RNA Green (Component A) is enough for one 96-well plate. Protect from light and avoid repeated freeze-thaw cycles. The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment. See figure 1 for details.

  3. For fixed cells: Fix cells with pure methanol for 1 minute at room temperature, then wash with PBS. Immerse cells in 1% Triton-100 for 2 minutes, then wash with PBS twice. Incubate cells with StrandBrite™ RNA Green (Component A) at the concentration of 1X in PBS at room temperature for 15 - 30 minutes. Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained. See figure 1 for details.

  4. (Optional) Wash the cells with PBS for 1 - 2 times, add 100 µL PBS to each well.

  5. Monitor fluorescence intensity with fluorescence microscope at Ex/Em = 490/520 nm (FITC channel).
Example data analysis and figures

Figure 1. Fluorescence images of RNA staining in HeLa cells. (A) Live cells were stained using Cell Navigator™ Live Cell RNA Imaging Kit (Green, Cat#22630) and counter-stained with Hoechst 33342 (Blue, Cat#17530). (B) Cells fixed in methanol were stained using the same kit. (C) After staining, fixed HeLa cells were incubated with 0.5 mg/mL RNase at 37 ºC for 1 hour. Image of RNase digest test indicates the high selectivity of StrandBrite™ RNA Green. The green fluorescence signal were measured using a fluorescence microscope with a FITC filter.
Disclaimer
AAT Bioquest provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please consult the Safety Data Sheet (SDS) provided for the product. Chemical analysis and/or reverse engineering of any kit or its components is strictly prohibited without written permission from AAT Bioquest. Please call 408-733-1055 or email info@aatbio.com if you have any questions.





Additional Documents

 
Safety Data Sheet (SDS)


Certificate of Analysis