Localization of RMST in CFs was detected using a lncRNA FISH Probe Mix kit (RiboBio, Guangzhou, China) following the manufacturer's protocol. Briefly, cells (2 × 104 cells/well) were seeded in a 4-chamber Lab-Tek II Chamber Slide and cultured in DMEM/F12 supplemented with 10% FBS for 24 h. Next, the cells were fixed with 4% paraformaldehyde at room temperature. Then, the fixed cells were treated with pre-cooled Triton X-100 (0.5% dissolved in 1× PBS). After washing with 1× PBS, the cells were incubated overnight with a 20 µM Cy3-conjugated FISH probe in the hybridization buffer (100 μL).
Lncrna fish probe mix kit
Manufactured by RiboBio
Sourced in China
The LncRNA FISH Probe Mix kit is a laboratory product designed to detect and visualize long non-coding RNA (lncRNA) molecules in cells through fluorescence in situ hybridization (FISH) techniques. The kit provides a set of pre-designed and pre-labeled probes that target specific lncRNA sequences, enabling researchers to study the localization and expression patterns of lncRNAs within biological samples.
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5 protocols using lncrna fish probe mix kit
1
Localizing lncRNA RMST in Cells
2
Fluorescence in situ Hybridization of lncRNA
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Fluorescence in situ hybridization (FISH) was performed using the lncRNA FISH Probe Mix kit (Ribobio, Guangzhou, China). MRC-5 cells were fixed in 4% paraformaldehyde (Solarbio, China) for 10 min and then added 1 ml precooling permeable solution (5 μl Triton-X + 1 ml PBS) for 5 min at 4 °C. Added 200 μl pre-hybridization buffer into each well then incubated at 37 °C for 30 min. Then discarded the pre-hybridization buffer and added 150 μl hybridization buffer with lncRNA FISH Probe Mix, hybridized overnight at 37 °C. Later washed with buffer I (4x SSC), buffer II (2x SSC) and buffer III (1x SSC) at 42 °C. The nuclei were stained with DAPI (Roche Molecular Biochemicals, Basel, Switzerland). The images were taken under the inverted fluorescence microscope (Olympus, IX73, Japan).
3
Localization of lncRNA ZNRD1-AS1 by FISH
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FISH assays were performed using the LncRNA FISH Probe Mix Kit (Guangzhou RiboBio Co., Ltd., Guangdong, China). Briefly, cells were fixed with 4% paraformaldehyde and treated with proteinase K and permeabilization solution. Following pre-hybridization at 37 °C for 30 min, cells were hybridized with fluorescently labeled ZNRD1-AS1 probe at 37℃ overnight. Nuclei were counterstained using DAPI and imaged using a confocal laser microscope.
4
Localization of lncRNA TP53TG1 by FISH
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To determine the cellular localization of lncRNA TP53TG1 in MRC-5, fluorescence in situ hybridization (FISH) was performed using the lncRNA FISH Probe Mix kit (Ribobio, Guangzhou, China). Briefly, cells were fixed in 4% paraformaldehyde (Solarbio, China) for 10 min at room temperature and then incubated in 1 mL pre-cooled permeable solution for 5 min at 4 °C. Then the cells were incubated with 200 μL pre-hybridization buffer for 30 min. Then the cells hybridized with the hybridization buffer containing lncRNA FISH Probe Mix at 37°C overnight. The next day cells were washed separately with 4 × SSC, 2 × SSC, and 1 × SSC at 42 °C for 5 min. Nuclei staining with DAPI (Roche, Basel, Switzerland). The images were taken under the inverted fluorescence microscope (Olympus, IX73, Japan).
5
RNA in situ Hybridization Using lncRNA FISH
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We conducted the RNA in situ hybridization through using lncRNA FISH Probe Mix Kit (RiboBio, China). We then seeded the cells in laser-confocal dishes, followed by 10 min fixation in 4% paraformaldehyde, and 5 min permeabilization with 0.5% Triton X-100, next we used PBS to wash them 3 times at room temperature. Cells were then soaked in prehybridization buffer at 37 ℃ for 0.5 h and incubated overnight at 37 ℃ with target probe hybridization buffer. We washed the cells with sodium citrate buffer and counterstained them with DAPI. Images were ultimately evaluated through confocal microscopy [25 (link)].
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