Flow cytometry was performed to assess the degree of podocyte apoptosis in vitro; double staining was conducted using the Annexin V-PE-7AAD Apoptosis Detection kit I (BD Biosciences) according to the manufacturer’s instructions. Samples were analyzed on a BD Accuri C6 flow cytometer (BD Biosciences) and cells in the upper- and lower-right quadrants were classified as apoptotic.
Annexin 5 pe 7 aad apoptosis detection kit 1
Manufactured by BD
Sourced in United States
The Annexin V-PE/7-AAD Apoptosis Detection Kit is a laboratory instrument used for the detection and analysis of apoptosis, a form of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and 7-AAD, a DNA-binding dye, to identify cells at different stages of the apoptotic process.
Automatically generated - may contain errors
Lab products found in correlation
Cell cycle analysis kit, by MultiSciences Biotech (2 mentions)
Facscalibur, by BD (2 mentions)
Accuri c6, by BD (1 mentions)
Naf s6776, by Merck Group (1 mentions)
Lymphocyte separation medium dkw33 r0100, by Dakewe (1 mentions)
Rabbit igg 7074p2, by Cell Signaling Technology (1 mentions)
Mouse igg 7076p2, by Cell Signaling Technology (1 mentions)
P perk sc 32577, by Santa Cruz Biotechnology (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
7 protocols using annexin 5 pe 7 aad apoptosis detection kit 1
1
Quantifying Podocyte Apoptosis via Flow Cytometry
2
Cell cycle and apoptosis analysis
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Cells were seeded at a density of 5 × 105 cells per well in 6-well plates. After treatment, cells were fixed in ice-cold 70 % ethanol. Cell cycle distribution was analyzed using the Cell Cycle Analysis Kit (MultiSciences, China) and apoptosis was detected using the Annexin V-PE/7-AAD Apoptosis Detection Kit I (BD BioSciences, USA) according to the manufacturer’s instruction. Stained cells were examined by FACSCalibur flow cytometry.
3
Apoptosis Assessment in Glioma Cells
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Apoptosis was quantified using Annexin V-PE 7AAD Apoptosis Detection kit I (BD) according to the manufacturer's instructions. U87 and GSC11 cells were incubated in standard culture conditions with and without drug treatment for 72 h before staining and subsequent flow cytometric analysis. We slso analyzed tumor tissue for evidence of apoptosis using a terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling kit per the manufacturer's instructions (Roche).
4
Endoplasmic Reticulum Stress Induction
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NaF (S6776) was purchased from Sigma Aldrich, UK. Lymphocyte separation medium (DKW33-R0100) were supplied by Dakewe Biotech Company, China. RIPA lysis buffer (P0013C), BCA Protein Assay Kit (P0012), Cell Counting Kit-8 (CCK-8) (C0038) and Annexin V-PE/7-AAD Apoptosis Detection Kit I (559763) were obtained from BD, USA. RPMI 1640 (11875119) and fetal bovine serum (16000044) were supplied by Gibco, UK. The mouse Bip (3177T), GRP94 (20292T), CHOP (2895P), ATF6 (65880S), p-JNK (4668T), p-eIF2a (3398P), ATF4 (11815S), cleaved caspase-12 (ab18766) antibodies, mouse IgG (7076P2) and rabbit IgG (7074P2) were obtained from Cell Signaling Technology, UK. p-PERK (sc-32577) were obtained from Santa Cruz Biotechnology, USA, p-IRE1 (ab48187) were obtained from Abcam, UK.
5
Apoptosis Detection in Glioma Cells
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Annexin V-PE/7-AAD Apoptosis detection Kit I (BD Bioscience, USA) was employed to determine the apoptosis of transfected glioma cells (U87 and U251). Cells were harvested after transfected with lentiviruses, washed twice with PBS, and then resuspended in 400 μl binding buffer. After the resuspension, cells were incubated with 5 μl Annexin V-PE and 10 μl 7-AAD solutions at room temperature for 10 min in the dark. The analysis was performed by flow cytometry (FACSCalibur; Becton Dickinson). For each sample, 10,000 cells were analyzed. All of the experiments were performed in triplicate. Transfection treatments include (1) anti-miR-21, (2) sh-CXCR4, (3) anti-miR-21 + sh-CXCR4, and (4) Lv-NC.
6
Apoptosis Analysis of Transfected Glioma Cells
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Annexin V-PE/7-AAD Apoptosis detection Kit I (BD Bioscience, USA) was employed to determine the apoptosis of transfected glioma cells (U87 and U251). Cells were harvested after transfected with lentivirus, washed twice with PBS, and then resuspended in 400 μl binding buffer.
After the resuspension, cells were incubated with 5 μl Annexin V-PE and 10 μl 7-AAD solutions at room temprature for 10 min in the dark. The analysis was performed by flow cytometry (FACSCalibur; Becton Dickinson). For each sample, 10,000 cells were analyzed. All of the experiments were performed in triplicate. Transfection treatments include: 1) anti-miR-21, 2) sh-CXCR4, 3) anti-miR-21 + sh-CXCR4, 4) Lv-NC.
After the resuspension, cells were incubated with 5 μl Annexin V-PE and 10 μl 7-AAD solutions at room temprature for 10 min in the dark. The analysis was performed by flow cytometry (FACSCalibur; Becton Dickinson). For each sample, 10,000 cells were analyzed. All of the experiments were performed in triplicate. Transfection treatments include: 1) anti-miR-21, 2) sh-CXCR4, 3) anti-miR-21 + sh-CXCR4, 4) Lv-NC.
7
Apoptosis and Cell Cycle Analysis
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Flow cytometry assay was performed using the Annexin V-PE/7-AAD Apoptosis Detection Kit I (BD Biosciences) according to the manufacturer's protocol. Briefly, 1 × 10 6 cells per well were collected and incubated with Annexin V/FITC and propidium iodide (PI) in the dark at room temperature for 15 minutes. Stained cells were examined with a FACScan flow cytometer (BD Biosciences), which was also used for cell cycle analysis. Before the test, cells were collected and fixed in 70% cold ethanol overnight at -20°C, followed by incubation with DNA staining solution and PI solution for 30 minutes in the dark at room temperature (Cell Cycle Analysis Kit, MultiSciences).
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