Annexin 5 propidium iodide double staining kit

Manufactured by BD

Sourced in United States

The Annexin V/propidium iodide (PI) double staining kit is a laboratory tool used to detect and quantify apoptosis and necrosis in cell samples. It utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a DNA-binding dye, to distinguish between viable, apoptotic, and necrotic cells through flow cytometry analysis.

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Lab products found in correlation

Cycletest plus dna reagent kit, by BD (1 mentions) Cyflow cube 6, by Sysmex (1 mentions) Fc500 flow cytometer, by Beckman Coulter (1 mentions) Facsverse flow cytometer, by BD (1 mentions)

7 protocols using annexin 5 propidium iodide double staining kit

Cell Cycle and Apoptosis Analysis in Cancer Cell Lines

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To analyze the influence of different treatments
on the cellular
activities of HCT116 and HepG2 cells, cell-cycle analysis was performed
using Cycletest PLUS DNA reagent kit (BD Cat no: 340242). The amount
of cell apoptosis was determined by Annexin V/propidium iodide double-staining
kit (BD Biosciences, Cat no: 556570). For the analysis, 1 × 10⁵ treated cells were trypsinized and washed with 1× PBS
and the suspended pellet was analyzed using a flow cytometer (CyFlow
Cube 6, SysmexPartec GmbH, Germany). Both analyses were performed
after 72 h culture duration.

Sengupta S., Khatua C, & Balla V.K. (2018). In Vitro Carcinoma Treatment Using Magnetic Nanocarriers under Ultrasound and Magnetic Fields. ACS Omega, 3(5), 5459-5469.

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Cell Apoptosis Measurement Protocol

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For measurement of cell apoptosis, briefly, cells were harvested 48 hours after
transfection, washed with phosphate-buffered saline (PBS), and fixed with 70% ethanol
overnight at 4°C. The cells were stained Annexin V/propidium iodide double-staining kit
(BD Biosciences, Waltham, Massachusetts) strictly according to the manufacturer’s
instruction and a FSCAN flow cytometer (FCM; BD Biosciences) was used to measure the cell
apoptosis.

Liu M.K., Ma T., Yu Y., Suo Y., Li K., Song S.C, & Zhang W. (2019). MiR-1/GOLPH3/Foxo1 Signaling Pathway Regulates Proliferation of Bladder Cancer. Technology in Cancer Research & Treatment, 18, 1533033819886897.

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Quantifying Apoptosis by Flow Cytometry

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Following transfection with Ep-3 siRNA or treatment with L-798106 for 48 h, the cells were harvested and stained with Annexin V/propidium iodide double staining kit (BD Biosciences) according to the manufacturer's protocol. Apoptotic cells were assessed by flow cytometry on an FC500 flow cytometer (Beckman Coulter, Inc., Brea, CA, USA).

Li L., Lv Y, & Yan D. (2018). Inhibition of Ep3 attenuates migration and promotes apoptosis of non-small cell lung cancer cells via suppression of TGF-β/Smad signaling. Oncology Letters, 16(5), 5645-5654.

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Annexin V/PI Assay for Apoptosis

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To study the effect of miR-29a on apoptosis, cells were stained with an Annexin V/propidium iodide (PI) double staining kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's protocol. Briefly, the cells were seeded at density of 3×10⁵ in 24-well plates, and were collected after 48 h of transfection, washed twice with cold PBS and resuspended in 1X binding buffer. Cells were stained with 5 µl Annexin V-FITC for 15 min and then with 5 µl PI for 10 min in the dark at room temperature. Apoptosis was measured by flow cytometry (BD Biosciences).

Gong H.L., Tao Y., Mao X.Z., Song D.Y., You D, & Ni J.D. (2019). MicroRNA-29a suppresses the invasion and migration of osteosarcoma cells by regulating the SOCS1/NF-κB signalling pathway through negatively targeting DNMT3B. International Journal of Molecular Medicine, 44(4), 1219-1232.

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Annexin V-FITC/Propidium Iodide Apoptosis Assay

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The Annexin V/propidium iodide (PI) double staining kit (BD Biosciences) was used to stain the cells according to the manufacturer's protocol. T-47D and SK-BR-3 cells were seeded in 24-well plates at a density of 3×10⁵ cells/well, transfected and collected at 48 h post-transfection. Subsequently, the cells were washed twice with cold PBS and resuscitated in 1X binding buffer. The cells were then stained in the dark with 5 µl Annexin V-FITC for 15 min and 5 µl PI for 10 min at room temperature. Apoptosis was detected using a BD FACSVerse flow cytometer (BD Biosciences) and analyzed using FlowJo v9.96 (FlowJo LLC).

Zhang J., Hu C., Hu D, & Fan Z. (2021). MicroRNA-1298-5p inhibits the tumorigenesis of breast cancer by targeting E2F1. Oncology Letters, 22(3), 660.

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Quantifying H/R-Induced Apoptosis

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H/R-induced apoptosis was monitored using the Annexin V-propidium iodide (PI) double-staining kit (BD Biosciences, San Jose, CA, USA) according to the instructions of the manufacturer, allowing quantification by flow cytometry (BD Biosciences). The proportion of apoptotic cells was defined as the percentage of Annexin V-positive cells relative to PI-positive cells.

Mao Y., Lee E., Yang X., Bae E.J., Jeon R, & Park B.H. (2022). Targeting p21-activated kinase 4 (PAK4) with pyrazolo[3,4-d]pyrimidine derivative SPA7012 attenuates hepatic ischaemia-reperfusion injury in mice. Journal of Enzyme Inhibition and Medicinal Chemistry, 37(1), 2133-2146.

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Metformin Induces Apoptosis

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Cells apoptosis was detected by Annexin V/Propidium Iodide (PI) double staining kit (BD Biosciences, USA) according to the manufacturer's instructions. After different concentrations of metformin (0, 50, 100, and 200 μM) for 48 h, cells were digested with trypsin, harvested and washed twice with PBS, resuspended in 200 μl binding buffer and labeled with 5 μl Annexin V and 5 μl PI in the dark. The apoptosis distribution was detected with flow cytometer, and the final ratio of apoptosis was calculated as the sum of early and late apoptosis.

Shen M., Yu H., Jin Y., Mo J., Sui J., Qian X, & Chen T. (2022). Metformin Facilitates Osteoblastic Differentiation and M2 Macrophage Polarization by PI3K/AKT/mTOR Pathway in Human Umbilical Cord Mesenchymal Stem Cells. Stem Cells International, 2022, 9498876.

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