Pi staining buffer

Manufactured by Merck Group

Sourced in United States, Germany

PI staining buffer is a laboratory solution used for DNA content analysis. It is designed to prepare cell samples for flow cytometry applications. The buffer contains propidium iodide, a fluorescent dye that binds to DNA, allowing the measurement of cellular DNA content.

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Lab products found in correlation

Accuri c6 flow cytometer, by BD (3 mentions) Modfit lt software, by Verity Software House (2 mentions) Facs caliber, by BD (1 mentions) Annexin 5 apc, by BioLegend (1 mentions) Facscan, by BD (1 mentions) Accuri c6 facscalibur flow cytometer, by BD (1 mentions) Click it edu proliferation assay kit, by Thermo Fisher Scientific (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions) Annexin 5 pi apoptosis detection kit, by BD (1 mentions) Propidium iodide, by Merck Group (1 mentions) Annexin 5 fitc, by Dojindo Laboratories (1 mentions) Propidium iodide, by Dojindo Laboratories (1 mentions)

Close spelling variants of this product

PI/RNase staining buffer

8 protocols using pi staining buffer

Cell Cycle Synchronization via Thymidine Double Block

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For thymidine double block [36 (link)], cells were seeded at 2 × 10⁵ cells per well in 6-well culture plates and treated with thymidine at 2 mM for 14 h. Cells were then washed, supplemented with normal media for 12 h, and treated with 2 mM thymidine for another 14 h. The cells were harvested at 2 h intervals up to 16 h and 26 h after release. Afterward, cells were harvested and fixed in 75% (v/v) cold ethanol at −20 °C for at least 2 h. The fixed cells were collected by centrifugation and resuspended in propidium iodide (PI) Staining Buffer (Sigma, St. Louis, MO, USA) to stain DNA and analyzed for DNA content on a flow cytometry (FACSCaliber; Becton Dickinson, Franklin Lakes, NJ, USA).

Lee J.H., Sung J.Y., Choi E.K., Yoon H.K., Kang B.R., Hong E.K., Park B.K., Kim Y.N., Rho S.B, & Yoon K. (2019). C/EBPβ Is a Transcriptional Regulator of Wee1 at the G2/M Phase of the Cell Cycle. Cells, 8(2), 145.

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Apoptosis and Cell Cycle Analysis

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Cells were treated with vehicle or indicated concentrations of enzastaurin and ibrutinib for 48 h for apoptosis and cell cycle analysis. For apoptosis assays, cells were stained with annexin V-APC (Biolegend, CA, USA) according to the protocol. For cell cycle assays, cells were stained with PI staining buffer (Sigma–Aldrich, Darmstadt, Germany) according to the manufacturer’s protocol. Finally, the labeled cells were analyzed using BD Accuri C6 flow cytometer (BD, Biosciences, San Jose, CA).

He Y., Li J., Ding N., Wang X., Deng L., Xie Y., Ying Z., Liu W., Ping L., Zhang C., Song Y, & Zhu J. (2019). Combination of Enzastaurin and Ibrutinib synergistically induces anti-tumor effects in diffuse large B cell lymphoma. Journal of Experimental & Clinical Cancer Research : CR, 38, 86.

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Honokiol Regulates Cell Cycle Markers

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Cells were incubated with either honokiol as indicated dosages or DMSO 0.01% for 24 or 48 hr. Cells were harvested and fixed with 70% ethanol overnight. After washing with PBS, cells were labelled with 500 μl PI staining buffer (Sigma‐Aldrich, St. Louis, MO) and incubated at room temperature in the dark for 30 min. DNA content was analysed using FACScan (Becton Dickinson, San Diego, CA) with ModFit LT 3.3 software. In addition, the cell cycle markers, such as cyclin D1, cyclin E, cdk4, cdk2, p21 and p27, were determined by Western blot with the antibodies (Cell Signaling, Danvers, MA, USA).

Huang K., Kuo C., Chen S., Lin C, & Lee Y. (2018). Honokiol inhibits in vitro and in vivo growth of oral squamous cell carcinoma through induction of apoptosis, cell cycle arrest and autophagy. Journal of Cellular and Molecular Medicine, 22(3), 1894-1908.

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Cell Cycle Analysis by PI Staining

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Cell cycle was analyzed by propidium iodide (PI) staining as described previously^{29 (link)}. In brief, cells were digested, washed with PBS, and fixed in the ice-cold 70% ethanol at −20 °C overnight. Then cells were stained with 40 μg/mL of PI staining buffer (Sigma) at room temperature for 30 min. Following analysis by the Accuri C6/FACSCalibur Flow Cytometer (BD Biosciences, USA), cell cycle data were processed with ModFit LT software (Verity Software House). All experiments were performed in triplicate.

Wang R., Li X., Sun C., Yu L., Hua D., Shi C., Wang Q., Rao C., Luo W., Jiang Z., Zhou X, & Yu S. (2021). The ATPase Pontin is a key cell cycle regulator by amplifying E2F1 transcription response in glioma. Cell Death & Disease, 12(2), 141.

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Cell Cycle Analysis of Murine Neurospheres

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Cell cycle analysis was performed using a Click-iT EdU proliferation assay kit (ThermoFisher, C10636), following manufacturer’s instructions. Briefly, primary murine neurospheres with fewer than 8 passages were dissociated with Accutase to generate single cell suspensions. 1 × 10⁵ cells were seeded in a well of a 12-well tissue culture plate and incubated overnight in the neurosphere medium. At the end of the 23rd hr, the medium was completely replaced with a fresh medium containing 10 μM Edu reagent. Cells were immediately returned to the incubator and grown for one more hour. The cells were lifted with Accutase, fixed with 4% PFA, and permeabilized with saponin, before the EdU was conjugated to a fluorophore with catalysis of a copper compound via a “click chemistry” reaction. The cells were then incubated in a PI staining buffer (200 μg/ml, Sigma, P4864) for 30 min at RT and analyzed on a BD LSR II flow cytometer.

Chen Z., Herting C.J., Ross J.L., Gabanic B., Vallcorba M.P., Szulzewsky F., Wojciechowicz M.L., Cimino P.J., Ezhilarasan R., Sulman E.P., Ying M., Ma’ayan A., Read R.D, & Hambardzumyan D. (2020). Genetic driver mutations introduced in identical cell-of-origin in murine glioblastoma reveal distinct immune landscapes but similar response to checkpoint blockade. Glia, 68(10), 2148-2166.

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Apoptosis and Cell Cycle Analysis

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For apoptosis and cell cycle analysis, cells were treated with indicated concentrations of HBI8000 and SHR2554. Then, cells were collected and treated with Annexin V/PI apoptosis detection kit (BD Biosciences, San Jose, CA, USA) and PI staining buffer (Sigma–Aldrich, Darmstadt, Germany) assay system according to the manufacturer’s instructions. Finally, all samples were analyzed by BD Accuri C6 flow cytometer (BD, Biosciences, San Jose, CA, USA).

Wang X., Wang D., Ding N., Mi L., Yu H., Wu M., Feng F., Hu L., Zhang Y., Zhong C., Ye Y., Li J., Fang W., Shi Y., Deng L., Ying Z., Song Y, & Zhu J. (2021). The Synergistic Anti-Tumor Activity of EZH2 Inhibitor SHR2554 and HDAC Inhibitor Chidamide through ORC1 Reduction of DNA Replication Process in Diffuse Large B Cell Lymphoma. Cancers, 13(17), 4249.

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Cell Cycle Analysis by Flow Cytometry

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Cells cultured in six-well plates were treated with FKBP14 shRNA or NC. After 48 h, cells were harvested and fixed overnight at 4°C in ice-cold 70% ethanol. The cells were then rehydrated and stained with propidium iodide (PI) staining buffer (Sigma-Aldrich, St. Louis, MO, USA) at room temperature for 30 min. Cells were then analyzed using flow cytometry (BD Biosciences, San Jose, CA, USA).

Lu M., Miao Y., Qi L., Bai M., Zhang J, & Feng Y. (2016). RNAi-Mediated Downregulation of FKBP14 Suppresses the Growth of Human Ovarian Cancer Cells. Oncology Research, 23(6), 267-274.

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Apoptosis and Cell Cycle Analysis of UNC2250

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Cells were treated with vehicle or indicated concentrations of UNC2250 for 12, 24, and 48 h for apoptosis assays or for 24 h for cell cycle analysis. For apoptosis assays, cells were stained with annexin-V-FITC and propidium iodide (PI) (Dojindo Laboratories, Kumamoto, Japan) according to the protocol. For cell cycle assays, cells were stained with PI staining buffer (Sigma–Aldrich, Darmstadt, Germany) as previously described [25 (link)]. All samples for apoptosis and cell cycle assays were analyzed by BD Accuri C6 flow cytometer (BD Biosciences, San Jose, CA, USA). The results of cell cycle assays were reanalyzed by ModFit LT software (Verity Software House, Topsham, ME, USA).

Shi C., Li X., Wang X., Ding N., Ping L., Shi Y., Mi L., Lai Y., Song Y, & Zhu J. (2018). The proto-oncogene Mer tyrosine kinase is a novel therapeutic target in mantle cell lymphoma. Journal of Hematology & Oncology, 11, 43.

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