Information about the subcellular localization of ZNF667-AS1 was gained with the help of a bioinformatics tool (http://lncatlas.crg.eu/ ). The subcellular localization of ZNF667-AS1 in Eca109 and Siha cells was verified by FISH performed using the Ribo™ lncRNA FISH Probe Mix(Red) (Guangzhou RiboBio Co., Ltd., Guangdong, China), with circular RNA TMEM87A and U2 snDNA as controls [20] (link). Five different visual fields were randomly selected for observation and photographing under a fluorescence microscope (Olympus Corporation, Tokyo, Japan)
Ribo lncrna fish probe mix red
Manufactured by RiboBio
Sourced in China
The RiboTM lncRNA FISH Probe Mix (Red) is a laboratory tool designed for the detection and visualization of long non-coding RNA (lncRNA) molecules in fixed cells or tissues. The product contains a mixture of fluorescently labeled probes that specifically bind to target lncRNA sequences, enabling their identification and localization within the sample.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microscope, by Olympus (50 mentions)
Ab104139, by Abcam (9 mentions)
Anti meg3 nucleotide probe, by Genecreate (2 mentions)
Inverted fluorescence microscope, by Olympus (2 mentions)
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
Hybridization solution, by RiboBio (1 mentions)
Fluorescence microscope, by Leica (1 mentions)
Confocal laser scanning microscope, by Leica (1 mentions)
Fish kit, by Roche (1 mentions)
Protease k, by Merck Group (1 mentions)
Microscopic fields, by Olympus (1 mentions)
67 protocols using ribo lncrna fish probe mix red
1
Subcellular Localization of ZNF667-AS1 via FISH
2
Subcellular Localization of LSAMP-AS1 in PCa Cells
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The subcellular localization of LSAMP-AS1 in PCa cells was examined by the FISH assay. The experiment was performed according to the instructions in the manual of Ribo™ lncRNA FISH Probe Mix (Red) (RiboBio, China). Briefly, the cells were seeded into the 24-well culture plate at the cell density of 6 × 104 cells/well and grew until the cell confluence reached 80%. The cells were then fixed with 1 mL of 4% paraformaldehyde at room temperature, followed by the treatments with proteinase K (2 μg/mL), glycine, and acetylation reagent respectively. Next, the cells were incubated with 250 μL of pre-hybridization solution at 42 °C for 1 h followed by overnight hybridization at 42 °C with 250 μL of pre-hybridization solution containing biotin-labeled antisense LSAMP-AS1 probe (300 ng/mL, Shanghai GeneChem Co., Ltd., Shanghai, China). After that, the cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (1: 800) diluted in phosphate buffered saline Tween-20 in a 24-well culture plate for 5 min. The cells were sealed with the anti-fade mounting medium. Five different visual fields were randomly selected for observation and imaging under the fluorescence microscope (Olympus, Tokyo, Japan) [24] (link).
3
Subcellular Localization of SNHG1 and miR-376a in HCC Cells
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Subcellular localization of SNHG1 and miR-376a in HCC cells was determined by FISH using Ribo lncRNA FISH Probe Mix (Red) (Ribo Biotechnology, Guangzhou, P.R. China). Cells were seeded at 6 × 104 cells/well in a 24-well culture plate. When cell confluence was about 80%, cells were washed with phosphate-buffered solution (PBS), and 1 mL of 4% paraformaldehyde was added. Cells were fixed at room temperature and permeated with 0.2% TX-100 at ambient temperature for 20 min. Cells were treated with Proteinase K (2 μg/mL), glycine, and acetylphthalating reagent. Pre-hybridization solution (250 μL) was added and incubated at 42°C for 1 h. Hybridization solution containing probe (250 μL, 300 ng/mL) was added and hybridized overnight at 42°C. After being washed three times with PBS buffer containing 0.1% Tween 20 (PBST), 4′-6-diamidino-2-phenylindole (1:800; Thermo Fisher Scientific, Waltham, MA, USA) was added to stain nucleus for 5 min. Cells were washed three times with PBST for 3 min each. Cells were mounted with an anti-fluorescence quencher. Five randomly selected fields were observed under a fluorescence microscope (Olympus, Tokyo, Japan).
4
Localization of BCYRN1 lncRNA via FISH
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BCYRN1 localization within cells was assessed via FISH using a Ribo™ lncRNA FISH probe Mix (Red) (Guangzhou RiboBio Co., Ltd.). Briefly, cells were added to coverslips in 24-well plates and were cultured overnight to 80% confluence, at which time they were fixed using 4% paraformaldehyde (1 mL) and treated with protease K, glycine, and ethyl phthalate (2 μg/mL). Cells were then treated for 1 h with 250 μL of a prehybridization solution at 42°C, followed by an additional 1 h incubation in hybridization buffer (250 μL) supplemented with the probe (300 ng/mL) at 42°C. DAPI (1 : 1000 in PBST) was then applied for 5 min to counterstain nuclei, and cells were mounted with an antifluorescence quencher and imaged via fluorescence microscope (Olympus Optical Co., Ltd), with five random fields of view per sample being examined.
5
MALAT1 Subcellular Localization by FISH
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MALAT1 subcellular localization was observed using FISH assay in strict accordance with the provided instructions of the Ribo™ lncRNA FISH Probe Mix (Red) (C10920, RiboBio Co., Ltd.). The cells (6×104 cells/well) were seeded into 24-well plates. Upon achieving 60–70% confluence, the cells were fixed using 4% paraformaldehyde, rinsed and permeabilized. The plates were sealed using the pre-hybridization solution. After elimination of the pre-hybridization solution, the cells were subjected to overnight hybridization at 37°C using the probe hybridization solution supplemented with anti-MALAT1 nucleotide (Wuhan Genecreate Bioengineering Co., Ltd.) in conditions devoid of light. Next, the cells were eluted, stained with DAPI, rinsed, and fixed using nail polish for observation under fluorescence microscopy (Olympus). Five different fields were selected, and the cells in these fields were observed and documented.
6
Subcellular Localization of lncRNA PAXIP1-AS1
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Subcellular localization of lncRNA PAXIP1-AS1 was identified using the FISH assay performed according to the standard instructions of Ribo™ lncRNA FISH Probe Mix (Red) (C10920, RiboBio Co., Ltd., Guangzhou, Guangdong, China). The cells were seeded (6 × 104 cells/well) in a 24-well culture plate and cultured until cell confluence reached 60–70%. The cells were then fixed in 4% paraformaldehyde, washed, and permeabilized. Pre-hybridization solution was employed to seal the plate. After the pre-hybridization solution was discarded, the cells were hybridized overnight at 37 °C with the probe hybridization solution containing anti-PAXIP1-AS1 nucleotide (Wuhan Genecreate Co., Ltd., Wuhan, Hubei, China) without light exposure. Following elution, the cells were stained with 4′,6-diamidino-2-phenylindole solution, rinsed, and mounted in nail polish for fluorescence microscopy (Olympus, Japan) and five different fields of view were selected for observation and photography.
7
Subcellular Localization of circ_000926 in A498 Cells
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Fluorescence in situ hybridization (FISH) was used to determine the subcellular localization of circ_000926 in A498 cells according to instructions provided by the RibolncRNA FISH Probe Mix (Red, RiboBio Co., Ltd., Guangzhou, Guangdong, China). A498 cells were inoculated in 24-well plates at a density of 3 Â 10 4 cells per well followed by 2-day culture. On reaching approximately 80% confluence, the cells were fixed in 1 mL of 4% paraformaldehyde for 10 minutes followed by treatment with 2 mg/mL of proteinase K, glycine, and acetylation reagent. Cells were then incubated with 250 mL of prehybridization solution and probed with 250 mL of 300 ng/mL of hybridization solution containing probe. The cells were then stained with DAPI dye (1:800) for 5 minutes. After the cells were sealed with an antifluorescence quenching agent, five visual fields were randomly selected and captured under a fluorescence microscope (Olympus Optical Co., Ltd., Tokyo, Japan).
8
Localization of circ_001653 via FISH
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FISH was utilized to identify the localization of circ_001653 in cells, according to the instructions of the Ribo lncRNA FISH Probe Mix (Red) (Guangzhou RiboBio, Guangzhou, Guangdong, China). The cells in logarithmic growth phase were seeded into the six-well culture plates at a density of 3 × 104 cells per well and cultured for 2 days until the confluence reached about 80%. The cells were fixed at room temperature with 1 mL of 4% paraformaldehyde for 10 min; treated with Proteinase K (2 μg/mL), glycine, and ethylphthalein; and incubated with the 250 μL of prehybridization solution at 42°C for 1 h. The cells were then hybridized with 250 μL of hybridization solution containing probe (300 ng/mL) at 42°C overnight. The next days, the cells were seeded into 24-well plates and stained for 5 min with DAPI prepared with PBS containing Tween 20 (PBST) and sealed by addition of an anti-fluorescence quenching agent. Five different visual fields were randomly selected and photographed under the fluorescence microscope (Olympus, Tokyo, Japan).
9
Subcellular Localization of HNF1A-AS1 by FISH
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FISH technique was applied for verifying the subcellular localization of HNF1A-AS1 in cells. Following the instructions of Ribo™lncRNA FISH Probe Mix (Red) (RiboBio Co., Ltd., Guangzhou, China), the cover plate was placed in a 24-well plate and cells were inoculated with 6 × 104 cells/well, so that the cells reached about 80% confluence. The glass was removed, and the cells were fixed with 4% paraformaldehyde (1 mL) at room temperature. Premixed solution (250 μL) was added after treated with protease K, glycine and acetylation reagent. Next, cells were incubated at 42 °C for 1 h. LncRNA HNF1A-AS1 (250 µL, 300 ng/mL) hybrid solution containing probe was added and crossed at 42 °C. After washed by phosphate-buffered saline with Tween (PBST, 0.01 M, pH 7.4) for 3 times, the nucleus were dyed with DAPI solution diluted by PBST (ab104139, 1:100, Abcam, Shanghai, China), then added to the 24-well culture plate and stained for 5 min. Finally, the cells were blocked with antifluorescence quenching agent, and a fluorescence microscope (Olympus, Tokyo, Japan) was adopted to observe and capture the images of cells.
10
Subcellular Localization of UCA1 by FISH
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The subcellular localization of UCA1 in cells was identified by FISH. According to the instructions of Ribo™ lncRNA FISH Probe Mix (Red) (RiboBio Co., Ltd, Guangdong, China), the specific methods were as follows: the slide was put into the 24-well culture plate, and the cells were seeded at 6 × 104 cells/well and grown to 80% confluence. The slide was taken out, the cells were fixed with 1 mL 4% paraformaldehyde after cleaning with PBS. After being treated with protease K, glycine and phthalylation reagent, the cells were added with 250 μL pre-hybrid solution and incubated at 42 ℃ for 1 h. The pre-hybrid solution was absorbed, cells were added with 250 μL UCA1 (300 ng/mL) hybrid solution containing probe and hybridized overnight at 42 ℃. The nucleus was stained with phosphate-buffered saline with Tween (PBST)-diluted DAPI (ab104139, 1:100, Abcam, Shanghai, China), added to the 24-well culture plate, and stained for 5 min. The cells were sealed with anti-fluorescence quenching agent, observed and photographed under a fluorescence microscope (Olympus Optical Co., Ltd, Tokyo, Japan).
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