Cells were digested with trypsin without EDTA (Gibco) and were washed twice with phosphate-buffered saline. The cells were treated with Annexin-V-FITC binding buffer, Annexin-V-FITC (Beyotime, China), and PI for 20 min at room temperature. Cell apoptosis was assessed by flow cytometric analysis using an FC500 flow cytometer (Beckman-Coulter). All experiments were performed in triplicate, and each data point represented the mean results of the triplicate experiments.
Annexin 5 fitc
Manufactured by Beyotime
Sourced in China, United States
Annexin V-FITC is a fluorescently labeled protein that binds to phosphatidylserine, a phospholipid that is exposed on the surface of apoptotic cells. It is commonly used in flow cytometry and fluorescence microscopy applications to detect and quantify apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Beyotime (85 mentions)
Facscalibur, by BD (28 mentions)
Facscalibur flow cytometer, by BD (14 mentions)
Flow cytometry, by BD (13 mentions)
Cellquest software, by BD (9 mentions)
Facscan, by BD (9 mentions)
Flow cytometry, by Beckman Coulter (8 mentions)
Trypsin, by Thermo Fisher Scientific (7 mentions)
Binding buffer, by Beyotime (7 mentions)
Mito tracker red cmxros, by Beyotime (6 mentions)
Facscan flow cytometer, by BD (6 mentions)
Annexin 5 fitc apoptosis detection kit, by Beyotime (5 mentions)
Annexin 5 binding buffer, by Beyotime (4 mentions)
Annexin 5 fitc binding buffer, by Beyotime (4 mentions)
Accuri c6 flow cytometer, by BD (4 mentions)
Propidum iodide, by Beyotime (3 mentions)
Novocyte, by Agilent Technologies (3 mentions)
Phalloidin tritc, by Merck Group (3 mentions)
Lsm 700 meta, by Zeiss (3 mentions)
Pi stain, by Beyotime (3 mentions)
273 protocols using annexin 5 fitc
1
Annexin-V-FITC Apoptosis Assay
2
Annexin V-FITC Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The transfected cells were digested with trypsin digestion solution without EDTA. The cell suspension was centrifuged at 1,000 g for 5 min. After discarding the supernatant, the cells were suspended in PBS and centrifuged again. The supernatant was then discarded. One hundred and ninety-five microliters of Annexin V-FITC (Beyotime, Hangzhou, China) binding solution was added to resuspend the cells, followed by 5 μl Annexin V-FITC and 10 μl PI (Beyotime, Hangzhou, China) staining solution. Flow cytometry was utilized to detect cell apoptosis.
3
Apoptosis Analysis of BMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After OGD exposure, BMSCs were digested and centrifuged. Then, cell pellets were incubated with 195‐μl Annexin V‐FITC binding solution with 10‐μl propidium iodide (PI) and 5‐μl Annexin V‐FITC (Beyotime Institute of Biotechnology) for 20 min. 4 × 105 cells in each group were assayed by flow cytometry within 1 h, and then, results were analyzed by CytExpert software (Beckman Coulter).
4
Annexin V-FITC Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse GC1-spg cells were prepared by trypsin (Gibco) digestion and centrifuged (L550, Xiangyi, Changsha, China) at 1000g for 5 min. Then, the cells were resuspended in PBS and counted. Aliquots of 100 000 resuspended cells were centrifuged at 1000g for 5 min. The supernatant was discarded and 195 μl Annexin V-FITC (Beyotime, Shanghai, China) was added. Cells were resuspended and treated with 5 μl of Annexin V-FITC, 10 μl of propidium iodide staining solution (Beyotime) was added, and cells were subjected to flow cytometry (Biosciences, Franklin Lakes, NJ, USA).
5
Apoptosis Evaluation in HUVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HUVECs (2 × 10 5 cells/well) were seeded onto 6-well plates. After the specified treatment, the cells were digested and collected by centrifugation at 1000 rpm for 5 min. The cells were washed twice in 1 ml PBS, and then gently resuspended by adding 195 μl of Annexin V-FITC (Beyotime C1056-1, China) binding solution. Thereafter, another 5 μl of Annexin V-FITC, 10 μl of propidium iodide staining solution (Beyotime C1056-2, China) was added. The cell suspension was incubated at the room temperature for 20 min in the dark. Finally, apoptosis in each group was immediately quantified though the flow cytometer (BD Biosciences, USA). FlowJo software (Ashland, OR) was sued for data analysis. All apoptosis assays were performed in duplicates or triplicates and all experiments were repeated at least three times.
6
Apoptosis Detection by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A flow cytometer (BD Biosciences) was used to detect the proportion of apoptotic cells. Treated cells were digested, resuspended and counted. Resuspended cells (5,000,000-10,000,000) were centrifuged at 1,000 x g for 5 min to obtain the cell precipitants. Subsequently, the cells were resuspended gently with 195 µl binding buffer for Annexin V-fluorescein isothiocyanate (FITC) staining. Next, following incubation with 5 µl Annexin V-FITC (Beyotime Institute of Biotechnology) for 15 min in the dark at 4˚C, 5 µl propidium iodide (PI) was added and cells were incubated for a further 5 min at 4˚C. Simultaneously, a tube without Annexin V-FITC or PI was used as a negative control. Subsequently, the cell apoptosis rates were determined by flow cytometry (FCM) and analyzed using BD Accuri™ C6 Software (Version 1.0.264.21; BD Biosciences).
Statistical analysis. All the statistical analyses were performed using GraphPad Prism 7.0 software (GraphPad Software, Inc., La Jolla, CA, USA). Student's t-tests were performed to assess the quantitative data compared between two groups, and one-way analysis of variance was used for multiple comparisons. All data were based on at least three independent experiments and values are presented as the mean ± standard deviation. P<0.05 was considered to indicate a statistically significant difference.
Statistical analysis. All the statistical analyses were performed using GraphPad Prism 7.0 software (GraphPad Software, Inc., La Jolla, CA, USA). Student's t-tests were performed to assess the quantitative data compared between two groups, and one-way analysis of variance was used for multiple comparisons. All data were based on at least three independent experiments and values are presented as the mean ± standard deviation. P<0.05 was considered to indicate a statistically significant difference.
7
Apoptosis and Autophagy Assays in SUDHL4 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were collected after transfection of 48 h. 1*10 4 cells were resuspended in 195μl Annexin V-FITC binding solution and 5μl Annexin V-FITC was added (Beyotime, Nanjing, China). Then, 10μl propidium iodide staining solution was added. After 20min, the apoptotic cells were detected through ow cytometry.
Immuno uorescent (IF) staining SUDHL4 cells were xed using 4% paraformaldehyde at 4 °C for 15 min after plasmids transfection, followed by permeabilized treatment with 0.2% Triton X-100 at room temperature for 15 min. Primary antibody against LC3 was added to incubate with cells at 4 °C overnight. Subsequently, cells were incubated with secondary antibodies for 90min. DAPI was used to stain nuclear. The cells were observed under an inverted microscope (Olympus, Tokyo, Japan).
Nude mouse xenograft model SUDHL4 cells were cultured to logarithmic growth stage in vitro, digested by trypsin, washed using PBS and resuspended in RPMI 1640 medium. The cells (2.5Í10 7 cells/ml) were subcutaneously injected into the left back of nude mice. The size of the tumor was measured after subcutaneous tumor formation.
The study was approved by the ethics committee of Jiamusi University.
Immuno uorescent (IF) staining SUDHL4 cells were xed using 4% paraformaldehyde at 4 °C for 15 min after plasmids transfection, followed by permeabilized treatment with 0.2% Triton X-100 at room temperature for 15 min. Primary antibody against LC3 was added to incubate with cells at 4 °C overnight. Subsequently, cells were incubated with secondary antibodies for 90min. DAPI was used to stain nuclear. The cells were observed under an inverted microscope (Olympus, Tokyo, Japan).
Nude mouse xenograft model SUDHL4 cells were cultured to logarithmic growth stage in vitro, digested by trypsin, washed using PBS and resuspended in RPMI 1640 medium. The cells (2.5Í10 7 cells/ml) were subcutaneously injected into the left back of nude mice. The size of the tumor was measured after subcutaneous tumor formation.
The study was approved by the ethics committee of Jiamusi University.
8
Epoxybutenolide-induced Apoptosis in SCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SCs were plated in 6-well plates (8 × 104/well) for 24 h, after which fresh media with or without 1 nM EpoB was added for an additional 24 h. In some experiments, 3-methyladenine (3-MA; 0.5 mM), IGF-1 (100 ng/ml), or LY294002 (20 μM; MedChemExpress, Shanghai, China) was added to the cultures and then maintained in the same media for the entire experiment. Cells were then suspended in 195 μl Annexin V-FITC binding buffer to which 5 μl Annexin V-FITC and 10 μl PI (Beyotime, Shanghai, China) was added. Following a 20 min staining period, flow cytometry (Beckman Instruments, Pasadena, CA, USA) was used to analyze samples in triplicate.
9
Quantifying Apoptosis by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Early and late apoptotic cells were assessed using flow cytometry. Cells were seeded into six-well plates at a density of 1x105 cells/well. Following transfection and treatment, apoptotic cells were analyzed. Briefly, cells in each group were collected and resuspended with 195 µl Annexin V-FITC (Beyotime Institute of Biotechnology). Cells (1x105) were subsequently treated with 5 µl Annexin V-FITC and 10 µl propidium iodide (both purchased from Beyotime Institute of Biotechnology) for 15 min at room temperature. Apoptotic cells were subsequently detected by flow cytometer (NovoCyte; ACEA Bioscience, Inc.) and analyzed by NovoExpress (version 1.2.5; ACEA Biosciences, Inc.).
10
Annexin V-FITC Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SUDHL4 cellswere collected after 48 h of transfection. 1*104 cells were resuspended in 195 µl of Annexin V-FITC binding solution with added 5 µl of Annexin V- FITC (Beyotime, Nanjing, China). Then, 10 µl of propidium iodide staining solution was added to incubate the cells for 20 min. Lastly, the apoptotic cells were detected by a flow cytometer.
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!