Pi solution

Manufactured by BD

Sourced in United States

PI solution is a laboratory reagent used in various analytical techniques. It serves as a buffer solution, maintaining a specific pH range to support various experimental protocols. The core function of PI solution is to provide a controlled chemical environment for sample analysis and processing.

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PI staining solution (38 citations) PI/RNase Staining Buffer Solution (32 citations) PI/RNase staining solution (29 citations) PI/RNase solution (17 citations) Annexin V-FITC and PI solution (7 citations) Propidium iodide solution (PI, (6 citations) PI working solution (4 citations) FxCycle PI/RNase staining solution (3 citations) FxCycleTM PI/RNAse Solution (2 citations)

27 protocols using pi solution

Flow Cytometry Analysis of EGFP and Propidium Iodide Staining in SFV/EGFP and SIN/EGFP Infected Cells

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Cells were infected on 6-well plates with SFV/EGFP and SIN/EGFP virus particles at an MOI of 10 as described above (1 ml of virus-containing solution was used for the infection). The infected cells were harvested 24 h after infection. Detached cells were harvested from the cell medium by centrifugation, and attached cells were trypsinized. The collected cells (approximately 10⁶) were washed with PBS and resuspended in 1 ml of PBS. For propidium iodide (PI) staining, the cells were incubated with 10 μl of 50 μg ml⁻¹ PI solution (Becton Dickinson Biosciences, San Jose, California, USA) and immediately processed for FACS analysis. EGFP and PI fluorescence was measured using a FACSAria II (Becton Dickinson Biosciences, San Jose, California, USA). The FACS data were analyzed by BD FACSDiva 6.1.2 software. Uninfected cells were used as a negative control for both the PI and EGFP FACS analysis and contained approximately 1-2% PI-positive cells in 4 T1 culture.

Zajakina A., Vasilevska J., Zhulenkovs D., Skrastina D., Spaks A., Plotniece A, & Kozlovska T. (2014). High efficiency of alphaviral gene transfer in combination with 5-fluorouracil in a mouse mammary tumor model. BMC Cancer, 14, 460.

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Cell Cycle Analysis of Transfected BV-2 Cells

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BV-2 cells were collected 48 h after transfection of plasmids. The cells were added to pre-cooled (−20°C) 75% ethanol and mixed overnight at 4°C. The cells were washed with PBS and treated with RNase, followed by staining with PI solution (Becton, Dickinson and Company) and incubation at 37°C for 1 h. The cells labeled by fluorescent are detected by Flow Cytometer. Subsequently, the cell cycle was assayed using Modfit software.

Wang Y., Han R., Xu Z., Sun X., Zhou C., Han B., He S, & Cong H. (2021). Upregulation of lncRNA147410.3 in the Brain of Mice With Chronic Toxoplasma Infection Promoted Microglia Apoptosis by Regulating Hoxb3. Frontiers in Cellular Neuroscience, 15, 648047.

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Cell Cycle Analysis by Flow Cytometry

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Synchronized cells under logarithmic growth phase were collected 24 h post synchronization, washed once with PBS and fixed using ice cold 100% ethanol in PBS. The samples were kept at −20 °C for at least 24 h. The fixed cells were washed twice with cold PBS and incubated in 200 µL PBS in the presence of RNase (0.25 mg/mL, Thermo Scientific, Waltham, MA, USA) for 30 min at 37 °C. For DNA staining, the cells were washed once with PBS and stained with 500 µL PI solution (50 µM, Invitrogen, Waltham MA, USA) in PBS for 30 min at 37 °C. Subsequently, the supernatant containing the PI solution was removed and the stained cells were resuspended in 500 µL PBS and read in BD FACSCanto™ II (Becton Dickinson, Franklin Lakes, NJ, USA). The cell cycle analysis was conducted by fitting a univariate cell cycle model using the Watson pragmatic algorithm as implemented in FlowJo v10.8 (FlowJo LLC, Ashland, OR, USA). It should be noted that the cell cycle assay provides a snapshot of cell percentages in different phases and gives valuable biological insights into cell dynamics, which are not directly comparable to a high-resolution live-cell proliferation assay.

Basti A., Malhan D., Dumbani M., Dahlmann M., Stein U, & Relógio A. (2022). Core-Clock Genes Regulate Proliferation and Invasion via a Reciprocal Interplay with MACC1 in Colorectal Cancer Cells. Cancers, 14(14), 3458.

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Cell Cycle and Apoptosis Analysis

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The SW620 cells (3×10⁶ cells/ml) were seeded into six-well plates and treated with 0.25 mg/ml LBF solution for 24 h. Following treatment, the cells were harvested and washed twice with PBS. For the cell cycle analysis, the cells were fixed in 70% ethanol overnight at 4°C. The fixed cells were then stained with PI solution (Sigma-Aldrich), which contained RNase A, for 45 min in the dark and analyzed by flow cytometry. For the Annexin V/PI staining assay, the cells were stained with Annexin V and PI solution for 10 min in the dark and analyzed by flow cytometry (Becton-Dickinson and Company, Franklin Lakes, NJ, USA). The untreated cells were used as a negative control.

WAN Y., XIN Y., ZHANG C., WU D., DING D., TANG L., OWUSU L., BAI J, & LI W. (2014). Fermentation supernatants of Lactobacillus delbrueckii inhibit growth of human colon cancer cells and induce apoptosis through a caspase 3-dependent pathway. Oncology Letters, 7(5), 1738-1742.

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Cell Cycle Analysis of MCF-7 Cells Treated with Capsicum Chinense Extracts and Nanoparticles

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Flow cytometry was used to investigate how C. chinense stem extract and NPs affect the cell cycle distribution of MCF-7 cells [18 (link)]. The cells, initially seeded at a density of 2.5 × 10⁵ cells/mL, were cultured in 6-well plates for 24 h before exposure to C. chinense stem extract (100, 250, and 500 µg/mL) and NPs (250, 500, and 1000 µg/mL) for an additional 24 h incubation. Following trypsinization, the cells were washed with PBS buffer and then fixed with ice-cold 70% ethanol at −20 °C. Subsequently, the cells were treated with PI solution (BD Biosciences, CA, USA) for 30 min at 4 °C. Flow cytometry, with the aid of BD Accuri C6 Plus software, was employed to analyze the cell cycle phases, and the resulting fluorescent signals were presented as histograms. Gated cells were manually classified into their respective cell cycle stages.

Chittasupho C., Samee W., Na Takuathung M., Okonogi S., Nimkulrat S, & Athikomkulchai S. (2024). Clerodendrum chinense Stem Extract and Nanoparticles: Effects on Proliferation, Colony Formation, Apoptosis Induction, Cell Cycle Arrest, and Mitochondrial Membrane Potential in Human Breast Adenocarcinoma Breast Cancer Cells. International Journal of Molecular Sciences, 25(2), 978.

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Apoptosis Analysis of SW1990 Cells

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SW1990 cells were seeded in 6-well plates at a density of 1.0 × 10⁵ cells per well. After 24 h incubation, the cell medium was replaced with fresh medium containing 100 μL of the Au DENPs or the polyplex solutions in each well for another 24 h. Then cells were collected and re-suspended in 100 μL ice-cold binding buffer, followed by addition of 5 μL of Annexin V-FITC and 5 μL of PI solution (BD Biosciences, San Diego, CA). The samples were incubated for 15 min in the dark at room temperature before being subjected to flow cytometry analysis. The assay was performed 3 times for each sample. The cells treated with PBS under the same processes were used as control.

Lin L., Fan Y., Gao F., Jin L., Li D., Sun W., Li F., Qin P., Shi Q., Shi X, & Du L. (2018). UTMD-Promoted Co-Delivery of Gemcitabine and miR-21 Inhibitor by Dendrimer-Entrapped Gold Nanoparticles for Pancreatic Cancer Therapy. Theranostics, 8(7), 1923-1939.

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Cell Cycle Analysis by Flow Cytometry

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Cells were seeded in 6-well dishes (Corning Incorporated-Life Sciences; Durham, NC, USA). After indicated treatments, cells were harvested and fixed in 70% ethanol at 4 °C for 16 h. The cell suspensions (1 × 10⁶/sample) were washed with 1 × PBS, and then stained with 10 μL of PI solution (BD Biosciences; San Jose, CA, USA). All samples were incubated in the dark at room temperature for 30 min. The cell cycle distribution was analyzed using a BD FACSCanto™ flow cytometer (San Jose, CA, USA).

Chen J.C., Lee I.N., Huang C., Wu Y.P., Chung C.Y., Lee M.H., Lin M.H, & Yang J.T. (2019). Valproic acid-induced amphiregulin secretion confers resistance to temozolomide treatment in human glioma cells. BMC Cancer, 19, 756.

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Cell Cycle Analysis of Transfected HeLa Cells

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Transfected HeLa cells were trypsinized and washed with PBS before they were fixed in 70% ice-cold ethanol overnight at 4°C. Then they were washed again with PBS and incubated in PI solution (BD Pharmingen) for 30 minutes at room temperature. The samples were examined by flow cytometry on a BD Flow Cytometry (BD Bioscience). 1 × 10⁴ cells per sample were collected for each condition. Data were analyzed using ModFit LT.

Liu J., Ji X., Li Z., Yang X., Wang W, & Zhang X. (2016). G protein γ subunit 7 induces autophagy and inhibits cell division. Oncotarget, 7(17), 24832-24847.

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Annexin V-APC/PI Apoptosis Assay

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The cell apoptosis was measured by AnnexinV-APC/propidium iodide (PI) double staining: cells in each group were centrifuged at 1000 rpm and 4°C for 5 min and collected. Binding buffer was 4 times diluted by deionized water (4 mL binding buffer+12 mL deionized water). Subsequently, the cells were washed by precooled PBS twice, and centrifuged at 1000 rpm for 5 min, with supernatant discarded, the cells were resuspended by 250 μL binding buffer, and the cell concentration was adjusted to 1 × 10⁶ cells/well; cell suspension (5 mL) was added into 5 mL flow tubes, then was added with 5 μL Annexin V-APC and 5 μL PI solution (both from BD Biosciences, Franklin Lakes, NJ, USA) and incubated without light exposure for 15 min. PBS (400 μL) was added into the reaction tubes, which was then injected in flow cytometry, the results were analyzed by a computer.

Huang X., Qiao F, & Xue P. (2019). The protective role of microRNA-140-5p in synovial injury of rats with knee osteoarthritis via inactivating the TLR4/Myd88/NF-κB signaling pathway. Cell Cycle, 18(18), 2344-2358.

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Cell Cycle and Apoptosis Analysis by Flow Cytometry

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The cells were seeded in 60-mm dishes and cultured overnight, and then treated with or without UA (40 μmol/L) for 24, 48 and 72 h. After treatment, floating and attached cells were collected and stained, and flow cytometric analysis was performed using a flow cytometer (FACSCanto II, BD Biosciences; San Jose, CA, USA). Cell cycles were evaluated by PI staining (PI solution, Dojindo) and apoptosis was detected using the Annexin V Cell Apoptosis Detection Kit 1 (BD Biosciences) according to the manufacturer’s instructions. Camptothecin (Merck, Darmstadt, Germany) was used as a positive control for the apoptosis assay.

Sahashi H., Kato A., Yoshida M., Hayashi K., Naitoh I., Hori Y., Natsume M., Jinno N., Kachi K., Asano G., Toyohara T., Kito Y., Ammanamanchi S, & Kataoka H. (2022). Urolithin A targets the AKT/WNK1 axis to induce autophagy and exert anti-tumor effects in cholangiocarcinoma. Frontiers in Oncology, 12, 963314.

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