Neutrophils purified from septic patients and healthy donors were stained with anti‐CD63 (PE, BioLegend, 353010), anti‐CD35 (FITC, BD, 565330), TUNEL (V450, BD, 561425), Annexin V PE/7‐AAD (BD, 559763) and CM‐H2DCFH‐DA (FITC, Solarbio, D6470). The samples were analysed by flow cytometry (FACSCanto II, BD Bioscience). Neutrophils were preincubated with or without cytochalasin B (CytB) before stimulation with PMA and then stained with anti‐CD63 (PE, BioLegend, 353010), anti‐CD35 (FITC, BD, 565330), Annexin V PE/7‐AAD (BD, 559763) and CM‐H2DCFH‐DA (FITC, Solarbio, D6470). The samples were analysed by flow cytometry (FACSCanto II, BD) and anti‐CD44 (PE, CST, 8724).
Annexin 5 pe 7 aad
Manufactured by BD
Sourced in United States
Annexin V-PE/7-AAD is a fluorescent-labeled reagent used for the detection and quantification of apoptotic cells. Annexin V binds to phosphatidylserine exposed on the cell surface during apoptosis, and 7-AAD is a DNA-binding dye that can detect late-stage apoptotic or necrotic cells. This reagent is commonly used in flow cytometry applications to identify different stages of cell death.
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Lab products found in correlation
Facscan, by BD (5 mentions)
Facscalibur, by BD (4 mentions)
Flow cytometry, by BD (3 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Fluorescence activated cell sorting (facs), by Beckman Coulter (2 mentions)
Facscalibur platform, by BD (2 mentions)
Annexin 5 fitc pi, by BD (2 mentions)
Facscanto 2, by BD (1 mentions)
Cm h2dcfh da, by Solarbio (1 mentions)
Confocal microscopy, by Zeiss (1 mentions)
Accuri c6 plus flow cytometry, by BD (1 mentions)
Alexa fluor 647 labeling kit, by Thermo Fisher Scientific (1 mentions)
Mcl 1, by Cell Signaling Technology (1 mentions)
Accuri c6, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Cellquest pro software, by BD (1 mentions)
Cocl2, by Merck Group (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Protease inhibitors cocktail, by Merck Group (1 mentions)
Cell counting kit 8 kit, by Dojindo Laboratories (1 mentions)
46 protocols using annexin 5 pe 7 aad
1
Neutrophil Activation and Apoptosis in Sepsis
2
Apoptosis Measurement in Drug-Treated Cells
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Annexin V-PE/7-AAD (BD Biosciences, San Jose, CA, USA) and terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL, Promega Corporation) assays were used to measure the percentage of drug-induced apoptotic cell death. Cells were seeded in 60 mm culture dishes or six-well plates at a density of ~1×106 cells per well and treated with aspirin, sorafenib or a combination at the indicated concentration. Cells treated with DMEM were used as the control group. Floating and adherent cells were collected after 24, 48 and 72 h of treatment. The Annexin V-PE/7-AAD stained cells were analyzed using flow cytometry (BD Biosciences), while TUNEL assay were only performed on cells after 48 h of treatment and stained cells were analyzed by confocal microscopy (Carl Zeiss).
3
Annexin V-PE/7-AAD Cell Apoptosis Assay
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The cell apoptosis was determined by Annexin V-PE/7-AAD (Becton Dickinson Co, NJ, USA) according to the instructions of the manufacturer. Approximately, 1 × 105 H9c2 cells were placed on six-well plates. Followed by the two washes with PBS, the H9c2 cells were collected and analyzed using BD Accuri® C6 Plus Flow Cytometry (BD Bioscience, NJ, USA).
4
Oxidative Stress-Induced Apoptosis in H9c2 Cells
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H9c2 cells were cultured in 6-well plates (1.5 × 105 cells per well). Afterwards, H9c2 cells were pretreated with TI (1 μM) for 2 h and treated with t-BHP (150 μM) for 6 h. After collection, cells were stained with Annexin V-PE/7-AAD and were detected by flow cytometry (Becton–Dickinson, NJ, USA).
5
Labeling and Dilution of rITs
6
Production and Analysis of Immunotoxins
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The immunotoxins dsFv-PE38, also Moxetumomab pasudotox [38 (link)], dsFv-PE24, also LR [12 (link)], the dsFv-PE24(B), and the Fab-PE24(B) [14 (link)] were produced, as previously described. The immunotoxin Fab-PE24 was cloned by exchanging the dsFv of LR with the Fab of Fab-PE24(B). The resulting protein was produced following standard procedures [39 (link)].
Secondary western blot antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), primary antibodies (Actin, GAPDH, and PARP) from Cell Signaling (Danvers, MA, USA), flow cytometry antibodies, and Annexin V-PE/7-AAD from Becton Dickinson (Franklin Lakes, NJ, USA).
Secondary western blot antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), primary antibodies (Actin, GAPDH, and PARP) from Cell Signaling (Danvers, MA, USA), flow cytometry antibodies, and Annexin V-PE/7-AAD from Becton Dickinson (Franklin Lakes, NJ, USA).
7
Apoptosis Evaluation via Flow Cytometry
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Control or treated cells were collected and stained with Annexin V-PE/7-AAD (Becton Dickinson, Franklin Lakes, NJ, USA). Apoptotic cell death was measured by counting the ratio of the AV-phycoerythrin positive cells, as determined by flow cytometry (Beyotime Biotechnology LLC).
8
Quantifying Cell Apoptosis by FACS
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Cells were cultured for 48 h, and then were harvested, washed, and resuspended in pre-cold PBS. Apoptotic cells were identified by staining with Annexin V-PE/7-AAD (BD Pharmingen, Franklin Lakes, NJ, U.S.A.) according to the manufacturer’s instructions. The stained cells were analyzed by fluorescence activated cell sorting (FACS) (Beckman Coulter, Brea, CA, U.S.A.).
9
Apoptosis Detection by Flow Cytometry
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Cells were harvested and resuspended with binding buffer, then stained with Annexin V-PE/7-AAD (BD Bioscience, USA) according to the manufacturer’s instruction. The apoptotic cells were quantified by flow cytometry (Accuri C6, BD Bioscience, USA).
10
Quantifying Apoptosis by Flow Cytometry
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