Fxcycle pi rnase staining solution

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany, Japan

The FxCycle PI/RNase Staining Solution is a reagent used for the detection and quantification of cellular DNA content in flow cytometry applications. It contains propidium iodide (PI) as the DNA-binding fluorescent dye and RNase to remove cellular RNA, allowing for the selective staining of DNA.

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Lab products found in correlation

Facscanto 2, by BD (14 mentions) Facscalibur, by BD (10 mentions) Facscalibur flow cytometer, by BD (8 mentions) Accuri c6, by BD (7 mentions) Lsr 2 flow cytometer, by BD (7 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (7 mentions) Accuri c6 flow cytometer, by BD (6 mentions) Facscanto 2 flow cytometer, by BD (5 mentions) Fitc annexin 5 apoptosis detection kit, by BD (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Propidium iodide, by Thermo Fisher Scientific (4 mentions) Facscalibur flow cytometer, by Thermo Fisher Scientific (3 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (3 mentions) Fortessa, by BD (3 mentions) Click it plus edu alexa fluor 488 flow cytometry assay kit, by Thermo Fisher Scientific (3 mentions) Cytoflex flow cytometer, by Beckman Coulter (3 mentions) Lsr 2, by BD (3 mentions) Macsquant flow cytometer, by Miltenyi Biotec (3 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (3 mentions) Click it plus edu flow cytometry assay kit, by Thermo Fisher Scientific (3 mentions)

Spelling variants of this product

FxCycle PI/RNase (17 citations) PI staining solution (15 citations) FxCycle PI/RNase Solution (13 citations) PI/RNase solution (11 citations) PI/RNase Staining Solution (10 citations) RNAse solution (8 citations) PI/RNase (6 citations) FxCycle PI (4 citations) FxCycle PI/RNase Staining (3 citations) FxCycle PI/RNase Staining Solution solution (2 citations)

280 protocols using fxcycle pi rnase staining solution

Flow Cytometric Analysis of Intestinal Epithelial Cells

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Flow cytometry analysis of intestinal epithelial cells was performed using standard protocols11 (link)70 (link). Propidium iodide (PI) stainings were performed with the FxCycle PI/RNase Staining Solution (Molecular Probes) following the manufacturer's instructions. Briefly, the cells were fixed with 4% paraformaldehyde and prepared at 1 × 10⁶ cells per condition. The cells were pelleted, followed by the addition of 0.5 ml of PI/RNase Staining Solution. After a 15-min incubation period, the samples were run on a BD Accuri C6 flow cytometer.

de Jong P.R., Taniguchi K., Harris A.R., Bertin S., Takahashi N., Duong J., Campos A.D., Powis G., Corr M., Karin M, & Raz E. (2016). ERK5 signalling rescues intestinal epithelial turnover and tumour cell proliferation upon ERK1/2 abrogation. Nature Communications, 7, 11551.

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Cell Cycle Regulation by IL11Rα and Doxorubicin

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AN3CA cells were cultured in serum free medium for 24 h to synchronize populations into G0. Medium was replaced with complete media containing 10% FCS, and cells treated with IgG control, or IL11Rα Ab (1 μg/ml) doxorubicin (500 ng/ml), or Ab and doxorubicin. Cells were harvested after 24 h and fixed overnight in 70% ethanol. Cells were stained with FxCycle PI/RNase staining solution (Molecular Probes) and analyzed on a BDFACSCanto II flow cytometer. Cell cycle and apoptosis analysis and model fitting was performed with FlowJo (FlowJo LLC).

Winship A., Van Sinderen M., Rainczuk K, & Dimitriadis E. (2017). Therapeutically blocking interleukin-11 receptor-α enhances doxorubicin cytotoxicity in high grade type I endometrioid tumours. Oncotarget, 8(14), 22716-22729.

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Apoptosis and Cell Cycle Analysis of MIA PaCa-2 Cells

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3 × 10⁵ MIA PaCa-2 cells per well were grown in 6 wells plate for 24 h. After 24 h, the cells were treated with 50 µM of drugs. After 72 h post treatment, both live and dead cells were collected, washed twice with cold PBS and the pellets were suspended in 500 µL binding buffer containing 5 µL of annexin-V FITC and 1 µL of propidium iodide (100 µg/mL) (Alexa Fluor^® 488 annexin V Apoptosis Kit, Invitrogen). The mixture was incubated for 15 min in the dark at room temperature and analyzed by flow cytometry. For cell cycle analysis, after harvesting total cells, the pellets were suspended in 1 mL of cold 70% ethanol and incubated overnight at 4 °C for fixation. The next day, cells were washed twice with cold PBS and suspended in 500 µl FxCycle PI/RNase staining solution (Molecular Probes) and analyzed by FACS Calibur flow cytometer after incubated at 37 °C for 30 min in the dark.

Kumar V., Chaudhary A.K., Dong Y., Zhong H.A., Mondal G., Lin F., Kumar V, & Mahato R.I. (2017). Design, Synthesis and Biological Evaluation of novel Hedgehog Inhibitors for treating Pancreatic Cancer. Scientific Reports, 7, 1665.

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Comparative Cell Growth and Characterization

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Cells were imaged with an IncuCyte HD (Essen BioScience, Ann Arbor, MI, USA) every 6-12 h during culturing, recording cell confluency for growth curves. Alternatively, since DIP2C KO cells differed in size, growth curves were generated by collection and counting of cells at set time points. For cell size comparison, cell diameter data was collected from the Cedex cell counter (Roche Innovatis, Switzerland) at eight occasions for a total of >5000 cells/cell line. For colony formation analyses, 400 cells plated in triplicate in 6-well plates were stained with 5% methylene blue in methanol after 10 days and colonies quantified. The plating efficiency was calculated as the number of obtained colonies divided by the number of seeded cells. For cell cycle analysis, equal numbers of cells fixed in ice cold 70% ethanol were stained with FxCycle PI/RNase staining solution (Molecular Probes, Eugene, OR, USA) for 15 min at room temperature, washed once in PBS, and analysed using a FlowSight flow cytometer (Amnis, Merck Millipore, Darmstadt, Germany).

Larsson C., Ali M.A., Pandzic T., Lindroth A.M., He L, & Sjöblom T. (2017). Loss of DIP2C in RKO cells stimulates changes in DNA methylation and epithelial-mesenchymal transition. BMC Cancer, 17, 487.

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Apoptosis and Cell Cycle Analysis of HDMB03 Cells

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3 × 10⁵ HDMB03 cells were transfected with scrambled and antimiR-217 and were grown in a 6-well plate for 72 hr. After 72 hr, both live and dead cells were collected and washed with cold PBS. For apoptosis assay, the cell pellets were suspended in 500 μL binding buffer containing 5 μL annexin-V fluorescein isothiocyanate (FITC) and 1 μL propidium iodide (Alexa Fluor 488 Annexin V/Dead Cell Apoptosis Kit, Invitrogen). The mixture was incubated for 15 min in the dark and analyzed by flow cytometry. For cell-cycle analysis, the HDMB03 cell pellets were suspended in 70% ethanol and incubated overnight. The cells were washed with PBS and suspended in FxCycle PI/RNase staining solution (Molecular Probes) and analyzed with a FACS Calibur flow cytometer after incubation at 37°C for 30 min in the dark.

Kumar V., Kumar V., Chaudhary A.K., Coulter D.W., McGuire T, & Mahato R.I. (2018). Impact of miRNA-mRNA Profiling and Their Correlation on Medulloblastoma Tumorigenesis. Molecular Therapy. Nucleic Acids, 12, 490-503.

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Cell Cycle Analysis by Flow Cytometry

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Following incubation for 48 h, cells were harvested and fixed overnight in cold 75% ethanol at 4°C. Next, cells were washed again with pre-cooled PBS and resuspended in 400 µl FxCycle™ PI/RNase staining solution (Molecular Probes; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. The samples were incubated for 30 min at room temperature in the dark, and then analyzed using a CytomicsTM FC500 flow cytometer (Beckman Coulter, Inc.). Cell cycle distribution was calculated with ModFit LT 4.1 software (Becton Dickinson and Company).

Xie J.H., Lai Z.Q., Zheng X.H., Xian Y.F., Li Q., Ip S.P., Xie Y.L., Chen J.N., Su Z.R., Lin Z.X, & Yang X.B. (2019). Apoptosis induced by bruceine D in human non-small-cell lung cancer cells involves mitochondrial ROS-mediated death signaling. International Journal of Molecular Medicine, 44(6), 2015-2026.

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Cell Cycle Analysis by Flow Cytometry

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To determine cell cycle distribution, cells were stained using propidium iodide (PI) and explored by flow cytometry. Following a period of culture of 24 h, JJN3 and U266 cells (2 × 10⁵ cells/mL/well) were incubated with 20–80 µg/mL of 5,6 α/β-ECs for 24–72 h. Cells were next fixed in 70% EtOH at −20 °C for 1 h and incubated with 500 µL of FxCycle PI/RNase staining solution (F10797, Molecular Probes, ThermoFisher Scientific, Waltham, MA, USA) for 30 min. At least, 10⁴ events were gated for each experiment using a BD FACSCanto II (BD Biosciences, Franklin Lakes, NJ, USA). Data were analyzed using the BD FACSDiva 7 software (BD Biosciences, Franklin Lakes, NJ, USA).

Jaouadi O., Limam I., Abdelkarim M., Berred E., Chahbi A., Caillot M., Sola B, & Ben Aissa-Fennira F. (2021). 5,6-Epoxycholesterol Isomers Induce Oxiapoptophagy in Myeloma Cells. Cancers, 13(15), 3747.

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BrdU-based Cell Proliferation and Apoptosis Assays

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BrdU (Sigma-Aldrich) was injected to 1-month-old mice with 100 mg/kg
intraperitoneally daily for five consecutive days. Immunohistochemical detection
of BrdU positive cells was performed using BrdU antibody (1:1000, Proteintech,
Rosemont, IL, USA). IMACs were incubated with 10 µM BrdU (Sigma-Aldrich)
for 2 hours then stained with anti-BrdU (1:200) primary antibody. To assess
apoptotosis, TUNEL staining was performed using In Situ Cell
Death Detection Kit (Roche Applied Science, Indianapolis, IN, USA). In MTT
assay, IMACs were incubated in DMEM supplemented with 10% FCS, and cells were
analyzed on days 1–5. MTT (5 mg/ml) was added. After adding DMSO,
absorbance values were measured at 540 nm. For cell cycle analysis,
fluorescence-activated cell sorting (FACS) analysis was performed using FxCycle
PI/RNase Staining Solution (Molecular Probes, Eugene, OR, USA). Cells were
resuspended and analyzed on a BD LSR II Flow Cytometer (BD Biosciences, San
Diego, CA). The percentages of cells in G1, S and G2/M phase were
determined.

Matsuzaki T., Alvarez-Garcia O., Mokuda S., Nagira K., Olmer M., Gamini R., Miyata K., Akasaki Y., Su A.I., Asahara H, & Lotz M.K. (2018). FoxO transcription factors influence cartilage maturation, homeostasis and osteoarthritis pathogenesis by modulating autophagy and proteoglycan 4. Science translational medicine, 10(428), eaan0746.

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Assessing Gemcitabine Sensitivity in Pancreatic Cancer

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MIA PaCa-2R+lenti-hsamiR205 and MIA PaCa-2R+lenti-hsamiRScramble cells were treated with 500 nM GEM after 72 h of culture. After 72 h of post-treatment, floating and attached cells were harvested, washed with cold PBS and fixed in 1 mL of 70% ethanol overnight at -20 °C. The next day, cells were washe d and resuspended in 500 μl FxCycle PI/RNase staining solution (Molecular Probes) and analyzed by an FACSCalibur flow cytometer. For apoptosis analysis, harvested cells were resuspended in 500 μl of 1× Annexin V binding buffer, 5 μl of Annexin V-Cy5, 5 μl of PI (BioVision Inc.) and analyzed by an FACSCalibur flow cytometer.

Chaudhary A.K., Mondal G., Kumar V., Kattel K, & Mahato R.I. (2017). Chemosensitization and Inhibition of Pancreatic Cancer Stem Cell Proliferation by Overexpression of microRNA-205. Cancer letters, 402, 1-8.

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Cell Cycle Analysis of VDX-111 Treatment

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Cells were plated and allowed to attach overnight before replacing media with media containing 100 nM VDX-111, 1 μM VDX-111, or equivalent volume of ethanol. After 72 hours of treatment, 1 x 10⁶ cells were fixed with ice cold 70% ethanol, washed with PBS, and resuspended in 500 μL FXCycle PI/RNase Staining Solution (Molecular Probes, Eugene, OR) for 15 minutes. Cells were then assessed using a Beckman Coulter Gallios flow cytometer with the 488 nm laser and 620/30 nm filter. Results were visualized using FlowJo (BD, Franklin Lakes, NJ) software by first selecting singlets based on forward and side scatter, then creating a histogram with gates around areas of interest.

Farrell K.B., Das S., Nordeen S.K., Lambert J.R, & Thamm D.H. (2024). VDX-111 targets proliferative pathways in canine cancer cell lines. PLOS ONE, 19(5), e0303470.

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