Cell apoptosis was determined with the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection Kit (BD Biosciences, San Jose, CA, USA) in accordance with the instructions. Jurkat cells were transfected with negative control, siLINC00680, and siLINC00680, along with a miR-320a inhibitor, and were cultured in a six-well plate at 5% CO2 at 37°C and with saturated humidity for 48 h. Subsequently, the cells were collected and washed with phosphate-buffered saline. Then, the cells were stained with annexin V-FITC and PI in the dark and the stained cells were measured by flow cytometry and analyzed by CellQuest software. These experiments were repeated in triplicate.
Annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis detection kit
Manufactured by BD
Sourced in United States
The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit is a laboratory reagent used to identify and quantify apoptotic cells. It contains Annexin V-FITC, which binds to phosphatidylserine, and propidium iodide, which stains DNA. This allows for the differentiation of viable, early apoptotic, and late apoptotic/necrotic cells through flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Cellquest software, by BD (3 mentions)
Flow cytometry, by BD (3 mentions)
Facscan flow cytometer, by BD (2 mentions)
Software, by FlowJo (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Facscalibur system, by BD (2 mentions)
Accuri c6, by BD (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Facs lsr 2, by BD (1 mentions)
Tc10 cell counter, by Bio-Rad (1 mentions)
Facsdiva software v7, by BD (1 mentions)
C6 flow cytometer, by BD (1 mentions)
Rnase a, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Binding buffer, by Beckman Coulter (1 mentions)
Flow cytometry, by Beckman Coulter (1 mentions)
Cytexpert software, by Beckman Coulter (1 mentions)
Facscan, by BD (1 mentions)
Facsverse flow cytometer, by BD (1 mentions)
50 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis detection kit
1
Apoptosis Evaluation in Jurkat Cells
2
Oxaliplatin-Induced Apoptosis in CRC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SW480 and SW620 cells growing in the logarithmic phase were collected and seeded into 6-well plates. After cell attachment, 1000 µg/L of oxaliplatin was added into the SW480 cells and 300 µg/L of oxaliplatin into the SW620 cells for an additional 48-h culture. Apoptosis rate was analyzed by flow cytometry using an Annexin V‐fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit (BD Biosciences). In short, SW480 and SW620 cells were collected and then re‐suspended in 1× binding buffer (500 μl) containing Annexin V‐FITC (20 μl) and PI (10 μl) in the darkness for 15 min at room temperature. Subsequently, apoptotic cells were evaluated by a fluorescence-activated cell sorting analyzer (Becton Dickinson). The results were analyzed using FlowJo 7.6.1 software.
3
Detecting Apoptosis in Cultured Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An Annexin V-Fluorescein Isothiocyanate (FITC)/Propidium Iodide (PI) Apoptosis Detection Kit (BD Pharmingen, CA, USA) was used to assay the rate of apoptosis of cultured cells. Cells were collected by trypsinization 12 h after H/R. After washing twice with cold PBS, the cells were resuspended in 300 ml of 1× binding buffer and stained with 5 ml of Annexin V-FITC plus 5 ml of PI for 15 min in the dark. Fluorescence signals were detected using a FACScan Flow Cytometer (BD Biosciences, CA, USA). Each experiment was conducted independently three times.
4
Annexin V-FITC/PI Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was detected using the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit (BD Biosciences) according to the manufacturer’s instructions. After treated with 10 mM Ala or Glu for 48 h, cells (1 × 106/well) were collected, centrifuged, and resuspended in 500 μl of 1 × binding buffer. Annexin V-FITC (5 μl) and PI (5 μl) were then added to each tube. The tubes were incubated in the dark at room temperature for 20 min. Immediately after incubation, cell apoptosis was assessed on a flow cytometry (BD Biosciences). Representative images of experiments were shown. All experiments were repeated at least three times.
5
Annexin V-FITC/PI Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For apoptosis detection, Min6 cells were stimulated with AS-IV, UA, or wortmannin for the indicated time. Thereafter, the cells were washed, centrifuged, and resuspended. Cell apoptosis was evaluated using the annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's instructions. Briefly, cells were incubated with 5 μl annexin V-FITC and 5 μl PI for 15 min at room temperature in the dark. A BD FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) was employed to detect cell apoptosis (early apoptosis + late apoptosis). The flow cytometry results were analyzed using CellQuest software (version 5.1; BD Biosciences, Franklin Lakes, NJ, USA).
6
Apoptosis Detection Using Annexin V-FITC/PI Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was detected using an Annexin V fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's instruction. Briefly, the cells were plated in 6-well culture plates at a density of 1.5×105 cells/well. After treatment with 20 μg/ml BKFE and/or 100 μM PA for 24 h, the cells were trypsinized and collected together with floating dead cells. The cells were then washed twice with cold DPBS and resuspended in 1× binding buffer. Annexin V-FITC and PI were added to the tube and the cells were incubated for 15 min at room temperature in the dark. Next, the cells were analyzed using flow cytometry (FACS LSR II, BD Biosciences, CA, USA). Annexin V−/PI− population was regarded as normal healthy cells. Apoptotic cells were determined by the sum of early and late apoptotic cells and were indicated as a percentage of the total number of cells.
7
HO-1, EZH2, and E2F Inhibition in Cell Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HO-1 enhancer Hemin was purchased from Sigma (St Louis, MO, USA). HO-1 inhibitor Znpp IX was purchased from Cayman Chemical (Ann Arbor, MI, USA). EZH2 inhibitor JQEZ5 and E2F inhibitor HLM006474 were purchased from MCE (Shanghai, China). The drugs were dissolved in a small amount of DMSO and stored in − 20 °C. Before using, we used serum-free RPMI-1640 to dilute it. Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection Kit was purchased from BD (San Diego, CA, USA). qPCR primers such as HO-1 and EZH2 were synthesized by Sangon Biotech (Shanghai, China). Western-blot was probed with primary antibodies, including antibodies against HO-1, EZH2, pRB. Secondary antibodies were purchased from Li-Cor Corp (Lincoln, Nebraska, USA). The TRIzol of total RNA extracted from cells and mononuclear cells was purchased from Invitrogen (Carlsbad, CA, USA).
8
Apoptosis Quantification in Suspension
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were treated with 3-MA (5 mM) or CQ (50 nM) and then cultured in medium of pH 6.6 or pH 7.4. Then cells were plated onto poly-HEMA-coated plates. After 48 hours in suspension, apoptotic cells were quantified (percentage) using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit (BD). The cells were harvested and counted using the TC10 Cell Counter (Bio-Rad, USA).
9
Berberine-Induced Apoptosis Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Annexin V-fluorescein isothiocyanate (FITC)/ propidium iodide (PI) Apoptosis Detection Kit (BD Pharmingen; Franklin Lakes, NJ, USA) was used to detect apoptosis according to the manufacturer’s instructions. After treatment with berberine for 10 h with or without pre-treatment with 3MA, z-VAD, or siTFEB, the cells were collected and incubated with 5 μL of Annexin V-FITC and 10 μL of PI for 15 min at room temperature in dark. After filtration, the cells were analyzed by fluorescence-activated cell sorting Calibur system within 1 h. Annexin V+/PI- and Annexin V+/PI+ represented apoptotic cells in early and late phases, respectively. The data were analyzed using BD FACSDiva Software v7.0 (Becton-Dickinson, USA).
10
Cell Proliferation and Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 48 h post-transfection, an MTT assay was applied to evaluate cell proliferation. Cells (3×103 cells per well) were seeded in a 96-well plate, and were incubated for 48 h followed by staining with 20 µl MTT (5 g/l) at 37°C for 4 h. Subsequently, the supernatant was discarded and the precipitate was dissolved following the addition of 200 µl dimethyl sulfoxide. The optical density value of each sample was measured at 490 nm using a spectrophotometer. Experiments were performed in triplicate. Cell apoptosis was determined using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection kit (BD Biosciences, Franklin Lakes, NJ, USA) using Flow cytometry. In brief, 48 h following cell transfection, cells were rinsed with cold PBS. Cells (5×105 cells/well) were then labeled with Annexin V-FITC and propidium iodide, according to the manufacturer's protocol. Flow cytometry (BD Biosciences, Franklin Lakes. NJ, USA) was used for cell apoptosis analysis. WinMDI version 2.5 (Purdue University Cytometry Laboratories; www.cyto.purdue.edu/flowcyt/software/Catalog.htm ) was used for data analysis and each test was repeated 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!