For cell cycle analysis, after 48 h treatment with UA (20 μM) or/and DIM (50 μM), TE-8 and TE-12 cells were collected, rinsed twice with cold DPBS, and fixed with 75% ethanol for 2 h at –20°C. Fixed cells were re-suspended in DPBS and stained with propidium iodide (PI, Sigma) for 30 min. For FITC/Annexin V staining analysis, cells were stained using a FITC Annexin V Apoptosis Detection Kit II (Becton-Dickinson Biosciences) at 37°C for 30 min. Stained cells were examined by a FACStar flow cytometer (Becton-Dickinson) and analyzed with Becton-Dickinson software.
Fitc annexin 5 apoptosis detection kit 2
Manufactured by BD
Sourced in United States, Germany, Belgium
The FITC Annexin V Apoptosis Detection Kit II is a laboratory product designed for the detection and quantification of apoptosis in cell samples. The kit utilizes Annexin V, a calcium-dependent phospholipid-binding protein, conjugated with the fluorescent dye FITC, to identify cells undergoing apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (21 mentions)
Facscalibur flow cytometer, by BD (11 mentions)
Facscanto 2, by BD (9 mentions)
Facsdiva software, by BD (8 mentions)
Facscanto 2 flow cytometer, by BD (7 mentions)
Lsrii flow cytometer, by BD (6 mentions)
Accuri c6, by BD (6 mentions)
Propidium iodide, by Merck Group (5 mentions)
Apo direct kit, by BD (4 mentions)
Accuri c6 flow cytometer, by BD (4 mentions)
β actin, by Santa Cruz Biotechnology (4 mentions)
Facsaria 3, by BD (3 mentions)
Methanol, by Merck Group (3 mentions)
Ethidium bromide, by Merck Group (3 mentions)
Acridine orange, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Lsrfortessa, by BD (3 mentions)
Fluorescence activated cell sorting (facs), by BD (3 mentions)
Facsverse, by BD (3 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (3 mentions)
208 protocols using fitc annexin 5 apoptosis detection kit 2
1
Cell Cycle and Apoptosis Analysis
2
Apoptosis Evaluation via Annexin V
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was determined by staining the cells with FITC Annexin V Apoptosis Detection Kit II (Becton-Dickinson Biosciences, CA, USA). The cells were harvested after incubation with UA (0, 10, 25 or 50 µM) 48 h and stained with Annexin V-FITC for 30 min. at 37 °C. The cells analyzed using a FACStar flow cytometer (Becton-Dickinson, San Jose, CA, USA).
3
Flow Cytometry Analysis of Resveratrol-Induced Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The flow cytometry method was used to study the intensity of the apoptotic induction under the effect of resveratrol. The cells of the tested lines (EPP85-181P, EPP85-181RNOV, AsPC-1 and H6c7) were cultured in 25 cm2 flasks for 24 h (37 °C, 5% CO2). After this time, cells were treated with resveratrol at concentrations of 0, 25, 50 and 100 µM for 48 h (37 °C, 5% CO2). Subsequently, they were trypsinized and then centrifuged (1050 rpm, 5 min) in fresh culture medium. Cells were then rinsed twice in PBS solution and centrifuged afterwards (1050 rpm, 5 min). Cells were diluted to 1 × 106 cells/mL and stained with the FITC Annexin V Apoptosis Detection Kit II (Beckton Dickinson, Franklin Lakes, NJ, USA) according to the manufacturer’s instructions. Data from at least 10,000 events were collected for each sample. The results obtained were further analyzed with the FlowJo 10.5 software (FlowJo, Asham, OR, USA). The experiment was carried out in three independent laboratory replications.
4
Apoptosis Assay Kit Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The APO-DIRECT TUNEL kit and FITC Annexin V Apoptosis Detection Kit II were obtained from Becton–Dickinson (BD Pharmingen, San Diego, CA, USA). Thiazole orange (TO), hydroethidine (HE), 3,3′-dihexyloxacarbocyanine iodide [DiOC6(3)], carbonyl cyanide m-chlorophenylhydrazone (CCCP) were obtained from Sigma-Aldrich (St Louis, MO, USA). Albumax II was obtained from Gibco (Gran Island, NY, USA).
5
Luminacin-Induced Apoptosis Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quantitative analysis of apoptotic cell death caused by Luminacin was performed using the FITC Annexin V Apoptosis Detection kit II (Becton Dickinson, Franklin Lakes, NJ), following the manufacturer’s protocols. Briefly, cells were plated at 1×106 cells/well in a 6-well plate, incubated for 16 hours, and then treated with luminacin (0, 1, 2.5, 5, 10, or 20 μM) for 24 hours. The cells were harvested, washed with cold PBS, and subjected to Annexin V–FITC and PI staining in binding buffer at room temperature for 10 minutes in the dark. The stained cells were analyzed by fluorescence-activated cell sorting (FACS ARIA3, BD Biosciences, San Jose, CA) using WinMDI 2.9 software (Purdue University Cytometry Laboratories, West Lafayette, IN).
6
Establishing HL-7702 Cell I/R Injury Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To construct the HL-7702 I/R injury model, HL-7702 cells were cultured in glucose-free RPMI 1640 medium (Gibco BRL/Life Technologies Inc.) and incubated in an oxygen-free atmosphere (95% N2 and 5% CO2 at 37°C) for 12 hours. Then, cells were cultured in normal RPMI 1640 medium under normal atmosphere (95% air and 5% CO2 at 37°C) for 24 hours (according to previous experiments in the hepatic I/R injury model16 (link),17 (link)). The cell apoptosis rate was detected using the FITC Annexin V Apoptosis Detection Kit II (Becton, Dickinson Co., Franklin Lakes, NJ, USA) with propidium iodide (AV/PI) according to the manufacturer’s instructions to establish the HL-7702 I/R injury model.
7
Annexin V Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After treatment with UA (0, 10, 20, 30, 40, or 50 µM) for 48 h, the TE-8 and TE-12 cells were harvested for the detection of apoptosis cells using the FITC Annexin V Apoptosis Detection Kit II (Becton Dickinson Biosciences, CA, USA). The cells were stained with Annexin V-FITC for 30 min at 37 °C and detected using an FACStar flow cytometer.
8
Quantitative Analysis of Radiation-Induced Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A quantitative analysis of apoptotic cell death caused by radiation was performed using a FITC Annexin V Apoptosis Detection kit II (Becton Dickinson, Franklin Lakes, NJ, USA). Cells were stained using an Annexin V-FITC apoptosis detection kit, following the manufacturer's protocol. Briefly, HaCaT cells were plated at 3×105 cells/well in a 6-well plate, incubated for 24 h, and then treated with KR22332 (10 µg/mL) only, radiation only (8 Gy), or radiation (8 Gy) plus KR22332 (10 µg/mL) for 72 h. The cells were then harvested, washed with cold PBS, and subjected to Annexin V-FITC and PI staining in binding buffer at room temperature for 10 min in the dark. The stained cells were analyzed by fluorescence-activated cell sorting (FACS ARIA3; BD Biosciences, San Jose, CA, USA) using WinMDI 2.9 software.
9
Cell Proliferation and Apoptosis Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation was detected 36 hours after transfection using the Cell Counting Kit-8 (Sangon, China) according to the manufacturer’s instructions. The cell apoptosis rate was detected 36 hours after transfection using the FITC Annexin V Apoptosis Detection Kit II with propidium iodide (AV/PI) (Becton, Dickinson Co.) according to the manufacturer’s instructions. In addition, expression of apoptotic markers cleaved caspase 3 (C-Caspase 3) and B-cell lymphoma-2 (Bcl-2) were detected at 36 hours after transfection using western blot, according to the detailed process described later.
10
Annexin V Apoptosis Detection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect apoptosis, we used the FITC Annexin V Apoptosis Detection Kit II (Becton Dickinson, USA). Annexin V conjugated with FITC has high affinity to phosphatidylserine, which is translocated to the outer part of cellular membrane in the early steps of apoptosis. To distinguish apoptosis and necrosis, propidium iodide (PI) staining was performed. After 6 h of treatment with the test inhibitors and drugs, cells were prepared as described in the manufacturer instructions and analyzed with a LSRII flow cytometer (Becton Dickinson, USA). For each experiment, positive (cells exposed to 100 µM camptothecin), negative (untreated cells), and unstained control samples were prepared. It is known, that apoptosis can be measured after different times of exposure to compounds: from short (4-6 h), medium (24 h) to long (48 h). We decided to choose 6 h treatment with drugs due to high cytotoxic effect observed after treatment with drugs. The apoptosis ratio was defined as a percentage of apoptotic (FITC-Annexin V positive, PI negative) cells in the sample, while necrotic cells as the percentage of PI positive cells. All experiments were performed in triplicate.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!