Cell Navigator® Live Cell RNA Imaging Kit *Green Fluorescence*
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.
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add StrandBrite™ RNA Green working solution
- Incubate for 30 - 60 minutes
- 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.
SAMPLE EXPERIMENTAL PROTOCOL
- Culture cells to a density optimum for imaging according to your specific induction protocol (about 1 - 2 × 104 cells/well/96-well plate).
- 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.
- 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.
- (Optional) Wash the cells with PBS for 1 - 2 times, add 100 µL PBS to each well.
- Monitor fluorescence intensity with fluorescence microscope at Ex/Em = 490/520 nm (FITC channel).
Spectrum
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Citations
View all 2 citations: Citation Explorer
Fine-tuning of highly bright benzo [c, d] indole-oxazolopyridine cyanine dye for nucleolar RNA imaging in living cells
Authors: Togashi, Nao and Nagaoka, Masaaki and Higuchi, Kei and Yoshino, Yukina and Wu, Yawen and Sato, Yusuke and Nishizawa, Seiichi
Journal: Talanta Open (2024): 100308
Authors: Togashi, Nao and Nagaoka, Masaaki and Higuchi, Kei and Yoshino, Yukina and Wu, Yawen and Sato, Yusuke and Nishizawa, Seiichi
Journal: Talanta Open (2024): 100308
Poly (ADP-ribose) polymerase 1 (PARP1) inhibition promotes pulmonary metastasis of osteosarcoma by boosting ezrin phosphorylation
Authors: Li, Fangfei and Wu, Xiaoqiu and Fu, Xuekun and Liu, Jin and Song, Wangze and Xiao, Gary Guishan and Lu, Aiping and Zhang, Ge
Journal: International Journal of Biological Sciences (2022): 1238
Authors: Li, Fangfei and Wu, Xiaoqiu and Fu, Xuekun and Liu, Jin and Song, Wangze and Xiao, Gary Guishan and Lu, Aiping and Zhang, Ge
Journal: International Journal of Biological Sciences (2022): 1238
Page updated on October 8, 2024