Pi staining buffer

Manufactured by BD

Sourced in United States

PI) staining buffer is a laboratory reagent used in cell and tissue analysis. It is designed to prepare samples for propidium iodide (PI) staining, which is a common method for measuring DNA content and cell cycle distribution. The primary function of the PI) staining buffer is to create an environment suitable for the PI dye to bind to cellular DNA, allowing for its detection and quantification using flow cytometry or other analytical techniques.

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Lab products found in correlation

Propidium iodide, by BD (2 mentions) Thymidine, by Merck Group (2 mentions) Facscalibur, by BD (2 mentions) Facsuit software, by BD (1 mentions) Facsverse, by BD (1 mentions) Facsuite software, by BD (1 mentions) Facs caliber, by BD (1 mentions) Modfit lt software version 3, by Verity Software House (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) 60 mm dishes, by Corning (1 mentions) Fluorescence activated cell sorting (facs), by BD (1 mentions)

Close spelling variants of this product

PI/RNase Staining Buffer BD Pharmingen PI/RNase staining buffer PI/RNase Staining Buffer kit PI)/ribonuclease staining buffer PI/RNase Staining Buffer Solution PI/RNA staining buffer Staining buffer

11 protocols using pi staining buffer

Cell Cycle Analysis of Melanoma Cells

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Melanoma cells were treated with the vehicle or the compounds at the indicated concentrations for 30 h or 45 h, then collected and fixed with 70% (w/v) ethanol at −20°C. Cells were washed twice with PBS and resuspended in PI Staining Buffer containing RNAse (Becton Dickinson, San Jose, CA, USA). Following incubation for 30 min at room temperature in the dark, cells were analyzed using a FACSVerse flow cytometer (Becton Dickinson). ModFit LT 3.0 software (Verify Software, Topsham, MN, USA) was used to calculate the percentages of cells in each cell cycle phase, and FACSuit software (Becton Dickinson) was used to calculate the percentages of dead cells in subG₁.

Sztiller-Sikorska M., Koprowska K., Majchrzak K., Hartman M, & Czyz M. (2014). Natural Compounds' Activity against Cancer Stem-Like or Fast-Cycling Melanoma Cells. PLoS ONE, 9(3), e90783.

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Cell Cycle Analysis of Melanoma Cells

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Melanoma cells were treated with PN for 24 h. Cells were collected and fixed with 70% (w/v) ethanol at −20°C. After washing with PBS, cells were resuspended in PI Staining Buffer containing RNase (Becton Dickinson). Following incubation for 30 min at room temperature, cells were analysed using a FACSVerse flow cytometer. ModFit LT 3.0 software (Verify Software) was used to calculate the percentages in each cell cycle phase and FACSuite software (BD Biosciences) to calculate the percentages of cells in subG₁.

Hartman M.L., Talar B., Sztiller-Sikorska M., Nejc D, & Czyz M. (2016). Parthenolide induces MITF-M downregulation and senescence in patient-derived MITF-Mhigh melanoma cell populations. Oncotarget, 7(8), 9026-9040.

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Quantifying Nuclear DNA in MCF-7 Cells

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The nuclear DNA contents of MCF-7/WT, MCF-7/ADR-1024 and MCF-7/ADR cells were determined using flow cytometry. At least 1×10⁵ cells were centrifuged and fixed with ethanol. Cells were resuspended in 500µL of PI-staining buffer (Becton Dickinson, San Jose, CA, USA) for thirty minutes before subjected to flow cytometer. The cells were analyzed immediately using a FACS Caliber (Becton Dickinson, San Jose, CA, USA). The fluorescence was measured with FL2 band-pass filter.

Tsou S.H., Chen T.M., Hsiao H.T, & Chen Y.H. (2015). A Critical Dose of Doxorubicin Is Required to Alter the Gene Expression Profiles in MCF-7 Cells Acquiring Multidrug Resistance. PLoS ONE, 10(1), e0116747.

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Annexin V-FITC and PI Apoptosis Assay

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After treatment, cells were collected by trypsin and then washed with PBS. On the day of analysis, cells were collected and washed twice with cold PBS. The cells were suspended in 5 µL Annexin V‐FITC and 10 µL propidium iodide (PI) staining buffer (BD) and incubated in the dark for >15 min. Stained cells were analyzed by FACS.

Zhong C., Zhu R., Jiang T., Tian S., Zhao X., Wan X., Jiang S., Chen Z., Gong R., He L., Yang J., Ye N, & Cheng Y. (2023). Design and Characterization of a Novel eEF2K Degrader with Potent Therapeutic Efficacy Against Triple‐Negative Breast Cancer. Advanced Science, 11(5), 2305035.

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Cell Cycle Analysis by Flow Cytometry

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Cells treated by GSK2126458 0.01μM for six or 24 hours, or without treatment were collected by centrifugation and resuspended at 1×10⁶ cells/mL in propidium iodide (PI) staining buffer (BD Bioscience, San Jose, CA). Cells were incubated at room temperature for 15 minutes. Cell-cycle histograms were generated after analysis of PI-stained cells by FACS with a BD FACSCalibur (BD Bioscience). For each culture, at least 1×10⁴ events were recorded. Histograms generated by FACS were analyzed by FACSDiv software (BD Bioscience).

Shukuya T., Yamada T., Koenig M.J., Xu J., Okimoto T., Li F., Amann J.M, & Carbone D.P. (2019). The effect of LKB1 activity on the sensitivity to PI3K/mTOR inhibition in non-small cell lung cancer. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 14(6), 1061-1076.

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Cell Cycle Analysis of Adenovirus-Infected Cells

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PAH PASMC or A549 cells were infected with serotype 5 adenoviruses carrying the indicated Mfn2 constructs for 24 hours at 100 pfu/cell. Following adenoviral infection, the cells were serum starved for 48 hours to achieve cell cycle synchronization and then stimulated with 10% FBS for 20 hours (for A549 cells) or 15% FBS for 24 hours (for PAH PASMC). Cells were harvested, resuspended in 300 μl 1× phosphate-buffered saline (PBS), and fixed with 70% ethanol (v/v) at −20°C. The fixed cells were washed with ice cold PBS and incubated with 500 μl of propidium iodide (PI)/staining buffer (BD Biosciences, Franklin Lakes, NJ, USA) at room temperature for 15 minutes. The samples were analyzed for DNA content using flow cytometry to detect PI binding to DNA using the SONY fluorescence-activated cell sorter (Sony SH-800, San Jose, CA, USA).

Dasgupta A., Chen K.H., Lima P.D., Mewburn J., Wu D., Al-Qazazi R., Jones O., Tian L., Potus F., Bonnet S, & Archer S.L. (2021). PINK1 induced phosphorylation of mitofusin 2 (Mfn2) at serine 442 causes its proteasomal degradation and promotes cell proliferation in lung cancer and pulmonary arterial hypertension. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 35(8), e21771.

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Cell Cycle and Apoptosis Analysis

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Cell distribution of ESCC cells was tested via propidium iodide (PI) staining followed by flow cytometry analysis. In brief, after fixed in 75% ethanol for 8 hours, cells were washed using phosphate-buffered saline (PBS) for 3 times (5 minutes per time). Next, cells were stained with PI staining buffer (BD Pharmingen, USA) for cell distribution analysis using a flow cytometry (FACSCalibur, BD Biosciences, USA). For cell apoptosis detection, treated ESCC cells were stained with PI and Annexin V-FITC for 15 minutes at room temperature, then used for cell apoptosis analysis using a flow cytometry.

Wang X., Gu M., Ju Y, & Zhou J. (2020). PIK3C3 Acts as a Tumor Suppressor in Esophageal Squamous Cell Carcinoma and Was Regulated by MiR-340-5p. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 26, e920642-1-e920642-11.

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Cell Cycle Analysis by Flow Cytometry

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Cell cycle distribution was determined by DNA flow cytometric analysis. Briefly, mock- or PRRSV-infected MARC-145 cells grown in 12-well plates at ~5 × 10⁵ cells/well were harvested by trypsinization, collected by centrifugation (200 g, 5 min), and washed twice with cold PBS. The cells were then fixed with cold 70% ethanol overnight at 4°C. After another washing step with cold PBS, the cell pellets were resuspended and stained with 500 μl of propidium iodide (PI) staining buffer (BD Biosciences, San Jose, CA, USA) containing 0.5 mg/ml RNase A for 15 min at room temperature in the dark. The cell cycle phases were determined by measuring DNA contents using a FACSCalibur flow cytometer (BD Biosciences), and the data were analyzed using the ModFit LT™ software Version 3.1 (Verity Software House, Topsham, ME, USA).

Wen X., Ge X., Zhou L., Zhang Y., Guo X, & Yang H. (2021). PRRSV Promotes MARC-145 Cells Entry Into S Phase of the Cell Cycle to Facilitate Viral Replication via Degradation of p21 by nsp11. Frontiers in Veterinary Science, 8, 642095.

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Cell Cycle Synchronization and Analysis

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MDCK cells were seeded at 25–30% confluence in 60 mm dishes (Corning). For cell synchronization, 2 mM thymidine (Sigma-Aldrich) was added, followed by incubation for 19 h at 37°C. After washing with PBS, fresh medium was added. The cells were incubated for 9 h at 37°C and exposed to 2 mM thymidine O/N for a second thymidine block. After cell synchronization, the medium was replaced with fresh medium with or without 12.5 μM FSK. Cells were collected at various times (2, 6, 12, or 24 h) and fixed in 75% ethyl alcohol for at least 2 h at –20°C. After washing, the cells were stained with propidium iodide (PI) staining buffer (BD Biosciences) for 15 min at room temperature (RT) and analyzed using a flow cytometer (Accurri; BD Biosciences, USA) and FlowJo software.

Lee S.Y., Lee J., Park H.L., Park Y.W., Kim H, & Nam J.H. (2023). The Adenylyl Cyclase Activator Forskolin Increases Influenza Virus Propagation in MDCK Cells by Regulating ERK1/2 Activity. Journal of Microbiology and Biotechnology, 33(12), 1576-1586.

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Cell Cycle Analysis by Flow Cytometry

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After treatment, cells were collected by trypsin and then washed with PBS. The cells were fixed with 70% ethanol for more than 18 h. On the day of analysis, cells were collected and washed twice with cold PBS. Suspend the cells in propidium iodide (PI) staining buffer (BD) and incubated in the dark for more than 15 min. PI-staining cells were analyzed by FACS (BD).

Guan Y., Jiang S., Ye W., Ren X., Wang X., Zhang Y., Yin M., Wang K., Tao Y., Yang J., Cao D, & Cheng Y. (2020). Combined treatment of mitoxantrone sensitizes breast cancer cells to rapalogs through blocking eEF-2K-mediated activation of Akt and autophagy. Cell Death & Disease, 11(11), 948.

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