The apoptosis of VCaP and PC3 cells was assessed using Annexin V/PI double staining kit (Invitrogen). After transfection, cells were reaped and re-suspended in Binding Buffer, and then double-stained in the dark for 15 min. After washing in PBS, the apoptotic cells were analyzed by flow cytometry (BD Biosciences, Franklin Lakes, NJ). The experiment was repeated at least three times.
Annexin 5 pi double staining kit
Manufactured by Thermo Fisher Scientific
The Annexin V/PI double staining kit is a laboratory tool used to detect and analyze apoptosis, or programmed cell death, in cell samples. The kit provides reagents and protocols for simultaneously staining cells with Annexin V and propidium iodide (PI), which bind to different cellular components to indicate the stage of the apoptotic process.
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Lab products found in correlation
Facscan, by BD (2 mentions)
Gapdh antibody, by Merck Group (1 mentions)
Rna extraction and reverse transcription kit, by Tiangen Biotech (1 mentions)
Sybr green universal qpcr master mix, by Tiangen Biotech (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Facscalibur, by BD (1 mentions)
Ow cytometer, by BD (1 mentions)
7 protocols using annexin 5 pi double staining kit
1
Quantifying Apoptosis in Prostate Cancer Cells
2
Evaluation of Anti-cancer Drugs in vitro
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Through our hospital pharmacy, we obtained VER from Shanghai Hefeng Pharmaceutical Company at 5 mg/2 mL, oxaliplatin (L-OHP) from Jiangsu Hengrui at 50 mg/ampul, doxorubicin hydrochloride (ADM) from Zhejiang Haizheng at 10 mg/ampul, 5-FU from Jiangsu Nantong Jinghua Pharmaceutical Company at 0.25 g/10 mL. Cell Counting Kit 8 (CCK-8) was provided by Japanese colleagues at the Chemical Institute. The Tiangen Company provided RNA extraction and reverse transcription kit and 2×SYBR Green Universal qPCR Master Mix. Mouse anti-human UCHL1 primary antibody was obtained from the Abcam company (USA). GAPDH antibody was purchased from Sigma; goat anti-mouse HRP-labeled secondary antibody was obtained from Guizhou Jinqiao Biological Company; and high-throughput sequencing was commissioned by Guangzhou Ruibo company. The Guangzhou Ruibo Company also provided siRNA for gene transfection; empty vector, and overexpression plasmid was purchased from the Origene Company, TrueORF GOLD model. The Beijing Beibo Company provided Annexin V-PI double staining kit; Lipofectamine 3000 was obtained from Invitrogen Company; Shanghai Shanjing Biotechnology Company conducted primer design and synthesis.
3
Analyzing Alcohol and Acetaldehyde Cytotoxicity
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HEMFs and esophageal epithelial cells treated with alcohol (0-200mM) and acetaldehyde (0-100uM) were analyzed by flow cytometry (BD FACSCalibur) after overnight incubation in KSFM or incubation with conditioned media from other cell type, with the markers Annexin V to detect cellular apoptosis and propidium iodide (PI) to detect necrotic or late apoptotic cells using the Annexin V/PI double staining kit (Invitrogen Catalog Number: V13242) according to manufacturer's instructions. Fluorescently-tagged Annexin V is a probe for membrane phosphatidylserine (PS) and PI enters cells with damaged membrane. Suspended cells were stained and immediately analyzed by flow cytometry. 10,000 cells were analyzed per measurement. Data was analyzed using Cyflogic V.1.2.1 software and the % of viable cells (PI-/Annexin V-), early apoptotic cells (PI-, Annexin V+), late apoptotic or necrotic cells (PI+, Annexin V+), and necrotic/unviable/dead cells (PI+/Annexin V-) were quantified (Rua
4
Quantifying Cell Apoptosis in Transfected Oral Cancers
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Cell apoptosis of transfected SCC-4 and SCC-9 cells was assayed employing the flow cytometer (BD Biosciences, Franklin Lakes, NJ), in the presence of Annexin V/PI double staining kit (Invitrogen). Cell samples were collected from 6-well plates via centrifugation, then stained in Binding Buffer and assayed with flow cytometry [18 (link)]. Each samples were assayed for more than triplicate.
5
Detecting Cell Apoptosis via Flow Cytometry
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Double Annexin V/PI staining kit (Invitrogen) was applied to estimate cell apoptosis. After 15 min of double-staining with binding buffer, cell samples were then determined with a flow cytometer (FACScan, BD Biosciences). The experiments were performed at least three times.
6
Apoptosis Quantification by Flow Cytometry
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Double Annexin V/PI staining kit (Invitrogen) was applied to estimate cell apoptosis. After 15 min of doublestaining with binding buffer, cell samples were then determined with a flow cytometer (FACScan, BD Biosciences). The experiments were performed at least three times.
7
Annexin V/PI Apoptosis Assay
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Apoptosis of transfected DU145 and LNCAP cells was estimated with application of double Annexin V/PI staining kit (Invitrogen). Cell samples were double-stained in 6-well plates with Binding Buffer for 15 min, then subjected to ow cytometer (BD Biosciences, Franklin Lakes, NJ).
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