Programmed cell death was analyzed by flow cytometry with Annexin-V-FITC/PI Apoptosis Assay Kit according to the manufacturer’s instruction (Thermo Fisher Scientific Waltham, Mass). PMNs were centrifuged, washed twice with ice-cold PBS, and resuspended in binding buffer. PMNs were then incubated with 5 μL Annexin-V-FITC reagent and 10 μL PI in the dark for 10 min, and then were analyzed by flow cytometry. The cells double-stained positive for Annexin-V and PI were considered to be programmed cell death. Cells were analyzed by BD LSRII flow cytometer (BD Biosciences, San Jose, Calif) and FlowJo software.
Annexin 5 fitc pi apoptosis assay kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Annexin V-FITC/PI apoptosis assay kit is a laboratory instrument designed to detect and quantify apoptosis, a programmed cell death process. The kit utilizes Annexin V-FITC and propidium iodide (PI) to identify cells undergoing early and late stages of apoptosis, respectively.
Automatically generated - may contain errors
Lab products found in correlation
Lsr 2, by BD (1 mentions)
Membrane protein extraction kit, by Thermo Fisher Scientific (1 mentions)
Hematoxylin and eosin h e, by Merck Group (1 mentions)
Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions)
Flowsight flow cytometry, by Merck Group (1 mentions)
Caspase 3 activity assay kit, by Beyotime (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
Facsaria fusion flow cytometer, by BD (1 mentions)
Dxp10, by Cytek (1 mentions)
Facs420, by BD (1 mentions)
Cellquest pro, by BD (1 mentions)
12 protocols using annexin 5 fitc pi apoptosis assay kit
1
Apoptosis Analysis by Flow Cytometry
2
Annexin V-FITC/PI Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Annexin V-FITC/PI Apoptosis Assay Kit (Thermo Fisher Scientific) was used to measure the effects of the different therapies on cell death. Briefly, the SW480 and HT29 cells were washed twice with ice cold PBS following the different treatments and re-suspended in 100 μl of 1× Annexin V (AV) binding buffer. Cell suspensions were then incubated in the dark for 15 min at room temperature, after the addition of AV-FITC (5 μl) and PI (1 μl). Before analysing with the Acea Novocyte 3000 flow cytometer, the samples were kept on ice after adding 400 μl of AV binding buffer. The percentages of live (non-stained), early (AV+/PI-) and late apoptotic (AV+/PI+), and dead (AV-/PI+) cells are shown as mean ± SD (n = 3).
3
Comprehensive Melanoma Cell Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cetyltrimethylammonium bromide (CTAB), dichloromethane (DCM), dimethyl sulfoxide (DMSO), tetraethylorthosilicate (TEOS), phosphate-buffered saline (PBS, pH 7.4; pH 6.5; pH 5.0), hematoxylin and eosin (H&E), methylthiazoletetrazolium (MTT), DTIC, fluorescein isothiocyanate (FITC) and Cy5.5 were purchased from Sigma-Aldrich. Annexin V-FITC/PI apoptosis assay kit, Membrane Protein Extraction Kit, the monoclonal antibody against CD47, E-cadherin and EpCAM were supplied by Thermo Fisher Scientific (USA). Melanoma cell line B16F10 cells were provided from American Type Culture Collection (ATCC, Manassas, VA, USA).
C57BL/6 mice (5–6 weeks old) were obtained by the Vital Laboratory Animal Center (Beijing, China). Mice were fed in our specific-pathogen-free facility at a temperature of 24°C. All animal experiments were approved by the Animal Ethical and Welfare Committee of Tianjin Medical University Cancer Institute and Hospital, and all animal studies were performed in compliance with the guidelines of the committee.
C57BL/6 mice (5–6 weeks old) were obtained by the Vital Laboratory Animal Center (Beijing, China). Mice were fed in our specific-pathogen-free facility at a temperature of 24°C. All animal experiments were approved by the Animal Ethical and Welfare Committee of Tianjin Medical University Cancer Institute and Hospital, and all animal studies were performed in compliance with the guidelines of the committee.
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
The Annexin V-FITC/PI Apoptosis Assay Kit (#V13245; Thermo Fisher Scientific) was used according to the manufacturer’s protocol. The cells were harvested after the different treatment protocols, washed twice with ice-cold PBS, and re-suspended in 100 µL of 1× Annexin V (AV) binding buffer. For cell staining, the mixture of AV-FITC (5 µL) and PI (1 µL) was added to each 100 µL of the SW480 and SW620 cell suspensions followed by incubation in the dark for 15 min at room temperature. Subsequently, the AV binding buffer (400 µL) was added, and the cells were placed on ice and immediately analysed using the NovoCyte 3000 flow cytometer. The experiments were processed in triplicate, and the data represent the percentage (mean ± SD) of the different apoptosis stages as follows: live (unstained), early (AV+/PI−) and late apoptotic (AV+/PI+), and dead (AV−/PI+) cells.
5
Annexin V-FITC/PI Flow Cytometry for Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was measured by an Annexin V-FITC/PI Apoptosis Assay Kit (#V13245; Thermo Fisher Scientific) as per the kit’s protocol. Following the different treatments, the SW480 and SW620 cells were collected, washed twice with cold-ice PBS, and re-suspended in 100 µl of 1× Annexin V (AV) binding buffer. A mixture of AV-FITC (5 µl) and PI (1 µl) was then added to each of the SW480 and SW620 cell suspensions followed by incubation in the dark for 15 min at room temperature for cell staining. Next, 400 µl of the AV binding buffer were added, and the cells were placed on ice and instantly analyzed with the NovoCyte 3000 flow cytometry. The experiments were conducted in triplicate and the data show the percentage (mean ± SD) of cells in the different apoptosis stages as follows: live (unstained), early (AV+/PI-) and late apoptotic (AV+/PI+), and dead (AV-/PI+) cells.
6
B16 Melanoma Cell Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
B16 cells (7.5 × 105 cells/well) were co-cultured with RAW 264.7 cells (1 × 106 cells/well) by using a 6-well transwell plate [49 (link)]. Briefly, cells were divided into four groups, including blank (only B16 cells), control (B16 cells co-cultured with RAW 264.7 cells), and 25 and 50 μg/mL of TFPS-treated (B16 cells co-cultured with RAW 264.7 cells) groups. After co-culturing for 24 h, B16 cells were collected and detected by Annexin V-FITC/PI apoptosis assay kit (Invitrogen, Carlsbad, CA, USA). Flowsight flow cytometry (Merck, Darmstadt, IN, USA) was used to analyze the apoptosis rate of B16 cells. Caspase-3 activity in B16 cells was performed by the Caspase 3 Activity Assay Kit (Beyotime, Shanghai, China).
7
Evaluating Cell Viability and Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Viability and apoptosis of MCF-7 and BT549 cells after various transfections were evaluated using the Cell Counting Kit-8 (CCK-8, Dojindo, Kumamoto, Japan) and Annexin V-FITC/PI Apoptosis Assay Kit (Invitrogen), respectively, referring to the recommendation of producers. Cell viability was proportional to the absorbance, which was detected at 450 nm using an iMark microplate reader (Bio-Rad, Munich, Germany). The FACScan flow cytometer (BD Biosciences, Le Pont de Claix, France) was used for the determination of cell apoptosis with Cell Quest software. Cell colony formation was evaluated using a standard colony formation assay as previously described.16 (link)
8
Annexin V-FITC Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An Annexin V-FITC/PI Apoptosis Assay Kit (Invitrogen Company, United States) was used to assess cell apoptosis. The cells were digested with trypsin, followed by washing twice with PBS. The 1 × 106 cells /mL cells were centrifuged for 5 min, and the supernatant discarded. Annexin-V-FITC labeling solution (20 μL) and PI reagent (20 μL) were successively added into 1 mL of buffer containing the cells, and incubated at room temperature in the dark for 5 min. A Beckman Coulter CytoFLEX LX Flow Cytometry System was employed to measure cell apoptosis. The experiment was repeated three times, and the data were averaged.
9
Quantifying Apoptosis in TZM-bl Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TZM-bl cells (1 × 106/well) were treated with TNF, AuNPs, and AuNP-TNF at 80, 420, and 500 µg/mL concentrations, respectively. According to the manufacturer's procedure (Annexin V-FITC/PI Apoptosis Assay kit; Invitrogen, New York, USA), staining of the treated cells was done for 10 min at RT in the dark with 5 µL PI for 5 min and 5 µL Annexin V-FITC. Cells without treatment served as a negative control, and cells treated with 10 µM Camtothectin (CPT), an inhibitor of DNA topoisomerase that induces Programmed Cell Death (PCD), served as a positive control. Using a FACSAria™ Fusion flow cytometer (Becton Dickenson, Franklin Lakes, USA), the amount of apoptosis induced by compounds in TZM-bl cells was quantified. In addition, apoptosis was evaluated using FlowJo software (version 10.0). The results were presented as the overall apoptosis rate (the early and late apoptotic cell population).
10
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 assay was performed Annexin V-FITC/PI apoptosis assay kit (Invitrogen, USA). Briefly, BT-474 and MCF-12A cells (5 × 105)/well were seeded into six-well plates. After 48 h miRNA transfection process, treated cells were harvested and washed twice with cold PBS. The treated cells were resuspended with 1 µl YO-PRO and 1 µl PI and incubated in ice for 30 min. Cell apoptosis rate was measured with flow cytometry (BD Accuri Plus, US).
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!