Cell Navigator® Live Cell RNA Imaging Kit Green Fluorescence

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Spectral properties

Excitation (nm)	509
Emission (nm)	527

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C)
UNSPSC	12352200

Overview SDS Protocol

Excitation (nm)

509

Emission (nm)

527

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.

Platform

Fluorescence microscope

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

Components

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

Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)	509
Emission (nm)	527

Images

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.

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

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