Pi working solution

Manufactured by Thermo Fisher Scientific

Sourced in United States

The PI working solution is a laboratory reagent used for the preparation of samples for various analytical procedures. It is a concentration of propidium iodide, a fluorescent dye, in a buffered solution. The PI working solution is commonly used in cell biology and flow cytometry applications to stain and identify cells.

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Lab products found in correlation

Annexin 5 alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Annexin binding buffer, by Thermo Fisher Scientific (1 mentions) Guava easycyte mini, by Merck Group (1 mentions) Facscanto 2, by BD (1 mentions) Annexin 5 fitc, by Thermo Fisher Scientific (1 mentions) Arthur image based cytometer, by NanoEnTek (1 mentions)

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PI solution (27 citations)

6 protocols using pi working solution

Quantifying TcdB Cytotoxicity in Cell Lines

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Confluent HeLa cells, 3T3 fibroblasts, and CHO cells grown in 96-well plates were intoxicated with 0.1 and 10 pM of TcdB from the different strains. The percentage of round cells in each well was evaluated every hour for a period of 12 h and then at 24 h.
For the cell death assay, confluent HeLa cells grown in 24-well plates were intoxicated with 100 pM of each toxin. Cytotoxicity was evaluated after 24 h of treatment. The cells were then harvested, washed in 1× PBS and resuspended in 100 µL 1× annexin binding buffer (Invitrogen, Waltham, MA, USA). Afterwards, 2 µL of Alexa Fluor 488 annexin V and 1 µL 100 mg/mL PI working solution (Invitrogen, Waltham, MA, USA) were added to the resuspended cells. After 15 min, the stained cells were analyzed by flow cytometry using a Guava easyCyte Mini (Merck Millipore, Burlington, MA, USA). The percentage of stained cells was determined with FLOWJO, LLC Data analysis software.

López-Ureña D., Orozco-Aguilar J., Chaves-Madrigal Y., Ramírez-Mata A., Villalobos-Jimenez A., Ost S., Quesada-Gómez C., Rodríguez C., Papatheodorou P, & Chaves-Olarte E. (2019). Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors. Toxins, 11(6), 348.

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Apoptosis Detection by Flow Cytometry

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Cells were collected and washed with 4 °C pre-cooled PBS, and then centrifuged at 10,000 r/min for 5 min at 4 °C. The cells was re-suspended in 1× Annexin binding buffer with the density of 10⁶/mL. 5 μL Alexa Fluor 488 Annexin V and 1 μL 100 μg/mL propidium iodide (PI) working solution (Invitrogen, Carlsbad, CA, USA) were added to each 100 μL cell suspension and incubated at room temperature for 15 min in dark and analyzed by flow cytometry. The data were analyzed using the workstation for flow cytometry (Beckman) and quantified by GraphPad Prism 5 software. One-way ANOVA and Tukey’s test were used for analysis of the statistical differences between test groups. P values < 0.05 were considered to be significantly different.

Zhang M.F., Luo X.Y., Zhang C., Wang C., Wu X.S., Xiang Q.P., Xu Y, & Zhang Y. (2022). Design, synthesis and pharmacological characterization of N-(3-ethylbenzo[d]isoxazol-5-yl) sulfonamide derivatives as BRD4 inhibitors against acute myeloid leukemia. Acta Pharmacologica Sinica, 43(10), 2735-2748.

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Annexin V-FITC/PI Cell Apoptosis Assay

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Cells were harvested and washed in ice cold PBS and resuspended in 1X Annexin-binding buffer at a density of 1x10⁶ cells/ml after centrifugation. FITC Annexin V (5 µl) and PI working solution (1 µl) (100 µg/ml) (Molecular Probes, Eugene, OR, USA) were added to 100µl of the cell suspension. After incubation for 30 minutes at room temperature, 400 µl of 1X Annexin‑binding buffer was added, mixed gently and the samples were kept on ice. The stained cells were immediately analyzed by flow cytometry (FACS Canto^TM II, BD BioSciences, San Jose, CA, USA).

Yin J., Li Z., Ye L., Birkin E., Li L., Xu R., Chen G., Ji J., Zhang Z., Jiang W.G, & Cui Y. (2020). EphB2 represents an independent prognostic marker in patients with gastric cancer and promotes tumour cell aggressiveness. Journal of Cancer, 11(10), 2778-2787.

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Annexin V-PI Apoptosis Assay

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The cells were seeded in 100-mm dishes at a density of 1,600,000 cells. After incubating overnight, the cells were treated with AOFE using the indicated concentrations. After 24 h, the cells were washed with PBS and treated with trypsin–EDTA solution. After harvesting, the cells were washed with cold PBS and resuspended in 1× annexin-binding buffer. Each 100 μL of cell suspension was incubated with 5 μL Alexa Fluor 488 annexin V and 1 μL 100 μg/mL PI working solution (Molecular Probes, OR, USA) for 15 min at room temperature. The prepared samples were analyzed using an Arthur image-based cytometer (NanoEnTek Inc., Seoul, Republic of Korea).

Cho E., Kim J., Jeong D.H, & Kim H.W. (2021). Anticancer properties of dried-pericarp water extracts of Camellia japonica L. fermented with Aspergillus oryzae through regulation of IGFBP-2/mTOR pathway. Scientific Reports, 11, 21527.

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Annexin V and PI Staining for Apoptosis Analysis

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We harvested and washed the cells, and resuspended the cells in 1 × annexin-binding buffer, added 5 μl Alexa Fluor 488 annexin V and 1 μl 100 μg/ml PI working solution (V13241, MOLECULAR PROBES, invitrogen, Eugene, OR 97402, USA) to each 100 μl of cell suspension and incubated the cells at room temperature for 15 min. After incubation, 400 μl 1 × annexin-binding buffer was added and gently mixed, while keeping the samples on ice. The stained cells were analyzed by flow cytometry, measuring the fluorescence emission at 530 and 570 nm using 488 nm excitation.

Xu Y.K., Ke Y., Wang B, & Lin J.H. (2015). The role of MCP-1-CCR2 ligand-receptor axis in chondrocyte degradation and disease progress in knee osteoarthritis. Biological Research, 48, 64.

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Annexin V and PI Cell Viability Assay

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The cells were seeded in 24-well plates at a density of 160,000 cells per well. After incubating overnight, the cells were treated with AOFE using the indicated concentrations. After 24 h, the cells were washed with PBS and incubated with 5 μL Alexa Fluor 488 annexin V and 1 μL 100 μg/mL PI working solution (Molecular Probes, OR, USA) for 15 min at room temperature. The cells were washed with PBS and fixed with 4% paraformaldehyde for 10 min. The cell images were photographed using a fluorescence microscope (Olympus IX71, Tokyo, Japan).

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