An Annexin V-FITC apoptosis detection kit (Elabscience, Houston, TX, USA) was used based on the manufacturer’s protocol. The cells were seeded in 6-well culture plates at a concentration of 3–4 × 105 cells/well and incubated overnight. NPS-1034 was added at 0, 10, and 40 μM and treated for 48 h. Treated cells were harvested, and the resulting pellets were resuspended in 100 ul of binding buffer and subsequently stained with 2 μL Annexin V-FITC and 2 μL PI. After incubating at room temperature for 15 min, the number of apoptotic cells was evaluated by flow cytometry (FACSCanto™ II Cell Analyzer; BD Biosciences, San Jose, CA, USA). For detection of the cell apoptotic rate, FlowJo software (BD Biosciences, San Jose, CA, USA) was used for formal analyses as described before [23 (link)]. All tests were performed in triplicate.
Annexin 5 fitc apoptosis detection kit
Manufactured by Elabscience
Sourced in China
The Annexin V-FITC Apoptosis Detection Kit is a laboratory product used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, a molecule that is translocated to the cell surface during apoptosis. The Annexin V is conjugated with the fluorescent dye FITC, allowing for the visualization and analysis of apoptotic cells.
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Lab products found in correlation
Cytoflex flow cytometry, by Beckman Coulter (2 mentions)
Cytexpert v2, by Beckman Coulter (2 mentions)
Facscalibur, by BD (2 mentions)
Facscanto 2 cell analyzer, by BD (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Aucubin, by Chengdu Must Bio-Technology (1 mentions)
Cell counting kit 8 cck 8, by Elabscience (1 mentions)
Pi rnase staining kit, by Dojindo Laboratories (1 mentions)
Penicillin streptomycin, by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Solarbio (1 mentions)
Cck 8, by Elabscience (1 mentions)
11 protocols using annexin 5 fitc apoptosis detection kit
1
Annexin V-FITC Apoptosis Detection Protocol
2
Apoptosis Analysis of Cancer Cell Lines
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A549, A549/DDP and NCI-H460 cells were seeded on 6-well plates and treated with different interventions for 48 h. Adherent cells were collected for apoptosis analysis after intervention. 5 µL Annexin V-FITC and 10 µL PI solution were provided by Annexin V-FITC Apoptosis Detection Kit (Elabscience, China) to stain the cells. CytoFLEX flow cytometry from Beckman Coulter, USA was used to detect apoptosis. CytExpert V2.3.0.84 software (Beckman Coulter, USA) was used to analyze apoptosis rates.
3
Quantifying ATDC5 Cell Apoptosis
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The cell apoptosis rate was detected using an annexin V‐FITC apoptosis detection Kit (Elabscience). Briefly, ATDC5 cells were seeded at a density of 1 × 106 cells/well in 6‐well plates overnight, then the cells were incubated with CM of various treatment groups for 48 h. After that, the dead and live cells were collected for centrifuging at 1000 g for 5 min and further incubated with Annexin V/PI (1:100 dye solution) for 15 min. The cells were washed with PBS and the cell death was assessed by FCM. Annexin V: λex = 488 nm, λem = 530 nm, PI: λex = 488 nm, λem = 630 nm. The data were quantified and then presented by FlowJo software.
4
Annexin V-FITC Apoptosis Assay
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The extent of apoptotic cell death in the different treatment groups was determined using the Annexin V-FITC Apoptosis Detection Kit (Elabscience Biotechnology Inc.). The cells were pre-incubated overnight in 6 cm dishes (1 × 106 cells). After various treatments, the cells were harvested and resuspended in 400 μL binding buffer containing Annexin V-FITC and propidium iodide (PI) according to the manufacturer’s instructions. The cell population in each group was measured using a BD Accuri C6 flow cytometer (BD Biosciences).
5
Annexin V-FITC Apoptosis Assay
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Cell apoptosis was assessed with the Annexin V-FITC apoptosis detection kit (Elabscience). Flow cytometry was performed with FACSCalibur (BD Biosciences) and all experiments were performed at least three times.
6
Aucubin Cytotoxicity Assay Protocol
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Aucubin with purity over 95% was supplied by Chengdu Must Bio-Technology Co. Ltd. (Sichuan, China). Roswell Park Memorial Institute (RPMI) 1640, fetal bovine serum (FBS), antibiotic-antimycotic solution, penicillin/streptomycin, phosphate-buffered saline (PBS), and 0.25% (w/v) trypsin/1 mM EDTA were procured by Gibco (Thermo Fisher Scientific, USA). Annexin V-FITC Apoptosis Detection Kit was purchased from Elabscience Biotechnology Co. Ltd. (Hubei, China).
7
Cytotoxic Effects of HA-SS-Geraniol in Prostate Cancer
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Geraniol was commercially bought from Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). F12K medium, fetal bovine serum (FBS), and penicillin-streptomycin (PS) were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China), Corning Inc. (Auckland, New Zealand), and Gibco, Thermo Fisher Scientific Inc., respectively. HA-SS-geraniol was provided by Dr. Duan’s lab at Henan University and synthesized as described previously.29 (link) The human PC-3 prostate cancer cell line was purchased from the Peking Union Medical College Cell Culture Center (1101HUM-PUMC000115) (Beijing, China). The Cell Counting Kit-8 (CCK-8) and Annexin V-FITC Apoptosis Detection Kit were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China). The Cellstain® Rhodamine 123 (Rh-123) Kit was procured from Sigma-Aldrich, Merck Ltd. The PI/RNase Staining Kit was purchased from Dojindo Molecular Technologies, Inc.
8
C3G Induces Apoptosis in A549 Cells
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A549 cells were seeded on 6-well plates and treated with different concentrations of C3G for 48 h. Adherent cells were collected for apoptosis analysis after intervention. 5 µL Annexin V-FITC and 10 µL PI solution were provided by Annexin V-FITC Apoptosis Detection Kit (Elabscience, China) to stain the cells. CytoFLEX flow cytometry from Beckman Coulter, USA, was used to detect apoptosis. CytExpert V2.3.0.84 software (Beckman Coulter, USA) was used to analyze apoptosis rates.
9
Apoptosis Evaluation of Fruit Extracts
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Flow cytometric measurement was performed using Annexin-V/7-AAD commercial kit to investigate the role of C. mas and B. vulgaris fruits extracts in the breast cancer cell line. After XTT, dilution ratios were created according to the IC50 values, and the extracts were added after the MCF-7 cells multiplied by 70%–80% by inoculating into sterile 6-well plates. Cells were incubated for 24 and 48 h. At the end of the period, the Annexin-V/7-AAD kit procedure was performed. Apoptosis analysis was performed using the “Elabscience® Annexin V-FITC Apoptosis Detection Kit”.
10
Culturing A549 Lung Cancer Cells
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The Human Non-Small Cell Lung Cancer Cells A549 were purchased from Procell Life Science and Technology (Wuhan, China), and maintained at 37 °C and 5% CO2 in DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin solution, respectively. The CCK-8 and Annexin V-FITC apoptosis detection Kit were obtained from Elabscience (Wuhan, China).
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