Annexin 5 fitc pi apoptosis detection kit

Manufactured by Abcam

Sourced in United States, United Kingdom

The Annexin V-FITC/PI Apoptosis Detection Kit is a laboratory product used to detect and quantify apoptosis in cell samples. It utilizes Annexin V conjugated to fluorescein isothiocyanate (FITC) and propidium iodide (PI) to identify and differentiate between early apoptotic, late apoptotic, and necrotic cells.

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Lab products found in correlation

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38 protocols using annexin 5 fitc pi apoptosis detection kit

Apoptosis Detection in H1650 Cancer Cells

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To estimate the pathway of H1650 cancer cell death, an Annexin V-FITC/PI apoptosis
detection Kit (BioVision, Mountain View, California) was used to detect the percentage of
apoptotic H1650 cells.^{20 (link)} Briefly, H1650 cancer cells were plated onto 6-well plate and transfected with
PS2ODNs when the cells reached 70% to 80% confluence. The mismatched sequence and
apoptosis inducer kit (Beyotime, Shanghai, China) were used as the negative and positive
controls, respectively. After incubation, the cells were harvested with 0.25% wt/vol
trypsin for apoptosis detection. Annexin V-FITC and PI were added for a 30-minute
incubation at 37°C in the dark to stain the harvested cells, followed by 2 washes with
phosphate-buffered saline. The stained cells in each group were detected by flow cytometry
(BD, New Jersey, USA) at an excitation wavelength of 488 nm and emission wavelengths of
525 and 625 nm.

Ge J.H., Zhu J.W., Fu H.Y., Shi W.B, & Zhang C.L. (2019). An Antisense Oligonucleotide Drug Targeting miR-21 Induces H1650 Apoptosis and Caspase Activation. Technology in Cancer Research & Treatment, 18, 1533033819892263.

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Detecting Apoptosis in PC12 Cells

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The annexin V-FITC/PI apoptosis detection kit (BioVision, Milpitas, CA, USA) was used to detect apoptotic cells according to the manufacturer's instructions. PC12 cells (1.5 × 10⁵ cells/well) were seeded into 6-well plates and cultured for 24 h at 37°C. Following incubation, cells were harvested, washed, and re-suspended in 300 μL PBS, containing 3 μL Annexin V-FITC and 3 μL PI for 30 min at room temperature. A total of 10,000 cells were collected and analyzed by flow cytometry. Experiments were performed in triplicate.

Li X., Li Y., Zhao J., Li L., Wang Y., Zhang Y., Li Y., Chen Y., Liu W, & Gao L. (2018). Administration of Ketamine Causes Autophagy and Apoptosis in the Rat Fetal Hippocampus and in PC12 Cells. Frontiers in Cellular Neuroscience, 12, 21.

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Apoptosis Detection in NSCLC Cell Lines

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Apoptotic cells were detected by an annexin V-FITC/PI apoptosis detection kit (BioVision, CA, USA) according to manufacturer's instruction. After incubation with navitoclax with or without pretreatment of Z-VAD-FMK (10 μM, 1 h) for 48 h, NCI-H1975 and NCI-H1975/OSIR cells were trypsinized, washed, and collected. Cells were then suspended in binding buffer with stained by Annexin-FITC and PI solution for 30 min. A total of 10,000 cells were collected and analyzed using a flow cytometer (FACS-Canto, BD Bioscience, USA).

Tang Z.H., Jiang X.M., Guo X., Fong C.M., Chen X, & Lu J.J. (2016). Characterization of osimertinib (AZD9291)-resistant non-small cell lung cancer NCI-H1975/OSIR cell line. Oncotarget, 7(49), 81598-81610.

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Flow Cytometric Analysis of Apoptosis

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Apoptotic cells were detected by an Annexin V-FITC/PI apoptosis detection kit (BioVision) according to manufacturer's instruction. MCF-7 cells and MCF-7/ADR cells were treated with different concentrations of EVO. After 48 h of incubation, cells were trypsinized and collected by centrifugation at 500 g/min for 5 min. After being washed twice with cold PBS and gently suspended in 100 µl binding buffer, cells were stained with 5 µl of Annexin-FITC and 10 µl of PI solution and incubated in the dark at room temperature for 15 min. Cell apoptosis was analyzed by a flow cytometer (BD Biosciences). All experiments were performed in triplicate.

Wang S., Wang L., Shi Z., Zhong Z., Chen M, & Wang Y. (2014). Evodiamine Synergizes with Doxorubicin in the Treatment of Chemoresistant Human Breast Cancer without Inhibiting P-Glycoprotein. PLoS ONE, 9(5), e97512.

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Apoptosis Detection by Flow Cytometry

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Differentiation among early apoptotic cells (annexin V +/PI −), late apoptotic cells (annexin V +/PI +), and necrotic cells (annexin V −/PI +) was analyzed by flow cytometry with a annexin V-FITC/PI apoptosis detection kit (cat # K101-25; BioVision Inc., Milpitas, CA, USA). After treating the harvested pellets with annexin V-FITC and propidium iodide, apoptosis detection and quantification were performed. The Attune NxT acoustic focusing cytometer (Thermo Fisher Scientific Inc., Waltham, MA, USA) was used to count the fluorescence of cells.

Sung H.C., Chang K.S., Chen S.T., Hsu S.Y., Lin Y.H., Hou C.P., Feng T.H., Tsui K.H, & Juang H.H. (2022). Metallothionein 2A with Antioxidant and Antitumor Activity Is Upregulated by Caffeic Acid Phenethyl Ester in Human Bladder Carcinoma Cells. Antioxidants, 11(8), 1509.

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Apoptosis Detection using Annexin V-FITC/PI

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Apoptotic cells were detected using an Annexin V-FITC/PI apoptosis detection kit (BioVision), following the manufacturer’s instructions. In brief, C6/36 cells from each treatment were collected by centrifugation at 3000 rpm for 5 min. After being gently suspended in 500 μL of binding buffer, cells were incubated with a mixture containing 5 μL Annexin V-FITC and 5 μL propidium iodide (PI) at 28 °C for 10 min in the dark. The Annexin V-FITC/PI-bound cells were ultimately analyzed with a FACScan flow cytometer (BD Biosciences).

Hou J.N., Chen T.H., Chiang Y.H., Peng J.Y., Yang T.H., Cheng C.C., Sofiyatun E., Chiu C.H., Chiang-Ni C, & Chen W.J. (2017). PERK Signal-Modulated Protein Translation Promotes the Survivability of Dengue 2 Virus-Infected Mosquito Cells and Extends Viral Replication. Viruses, 9(9), 262.

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Quantifying Apoptosis and Necrosis by Flow Cytometry

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Annexin V-FITC/PI Apoptosis Detection Kit (BioVision, USA) was used for the flow cytometry assay to quantify the apoptotic and necrotic cells on Days 4 and 6 of culture. This test was followed as per the manufacturer’s instruction by flow cytometry (FACS Calibur; BD Biosciences, USA) using the FITC signal detector (FL1) and by PI staining using the phycoerythrin emission signal detector (FL2). Data were analyzed by recording 5,000 events with Cell Quest Pro software (BD Biosciences) (Figure S1A).

Das K., Madhusoodan A., Mili B., Kumar A., Saxena A., Kumar K., Sarkar M., Singh P., Srivastava S, & Bag S. (2017). Functionalized carbon nanotubes as suitable scaffold materials for proliferation and differentiation of canine mesenchymal stem cells. International Journal of Nanomedicine, 12, 3235-3252.

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Annexin V-FITC Apoptosis Assay

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The extent of apoptosis was determined by flow cytometry via Annexin V-FITC-PI apoptosis detection kit (Biovision). Briefly, SH-SY5Y cells and SH-SY5Y-ROP16 cells were harvested and rinsed twice with cold PBS (pH7.4) respectively before resuspended in 1×binding buffer at a concentration of 1×10⁶ cells/ml. 5 µl of Annexin V-FITC and 5 µl of propidium iodide were added to 500 µl of cell suspension and then were incubated for 15 mins at room temperature in darkness. The stained samples were analyzed by Flow cytometer system (FACS Calibur BD Flow Cytometer) and Cell Quest software (BD) within 1 hour.

Chang S., Shan X., Li X., Fan W., Zhang S.Q., Zhang J., Jiang N., Ma D, & Mao Z. (2015). Toxoplasma gondii Rhoptry Protein ROP16 Mediates Partially SH-SY5Y Cells Apoptosis and Cell Cycle Arrest by Directing Ser15/37 Phosphorylation of p53. International Journal of Biological Sciences, 11(10), 1215-1225.

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Quantifying Apoptosis by Flow Cytometry

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Flow cytometry was performed to quantify apoptotic cells using an Annexin V-FITC/PI Apoptosis Detection kit (BioVision, Inc., Milpitas, CA, USA). HK-2 cells were harvested and washed in ice-cold PBS twice and double-stained with Annexin V-FITC and PI at room temperature for 30 min in the dark. All samples were quantitatively analyzed using a FACSCalibur flow cytometer at 488 nm emission and 570 nm excitation (BD Biosciences, San Jose, CA, USA) and analyzed by CellQuest software (version 3.0; BD Biosciences).

Wang Y., Zhang W., Yu G., Liu Q, & Jin Y. (2018). Cytoprotective effect of aquaporin 1 against lipopolysaccharide-induced apoptosis and inflammation of renal epithelial HK-2 cells. Experimental and Therapeutic Medicine, 15(5), 4243-4252.

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Apoptosis analysis of cancer cell lines

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A549, A549/CR, H460 and H460/CR cells after exposure to PGG at a level of 25 μM for 48 h were labeled with an Annexin VFITC/PI apoptosis detection kit (Biovision Inc, Mountain View, CA, USA). Then, the numbers of apoptotic and necrotic cells were determined by the Cell Quest software of BD flow cytometer.

Kim J.H., Im E., Lee J., Lee H.J., Sim D.Y., Park J.E., Ahn C.H., Kwon H.H., Shim B.S., Kim B, & Kim S.H. (2022). Apoptotic and DNA Damage Effect of 1,2,3,4,6-Penta-O-galloyl-beta-D-glucose in Cisplatin-Resistant Non-Small Lung Cancer Cells via Phosphorylation of H2AX, CHK2 and p53. Cells, 11(8), 1343.

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